Muco-adhesive, controlled release formulations of levodopa and/or esters of levodopa and uses thereof

ABSTRACT

The invention provides a controlled release oral solid formulation comprising (a) a controlled release component comprising core comprising levodopa and/or an ester of levodopa or salts thereof, wherein the core is coated with a layer of a muco-adhesive polymer and externally coated with a layer of an enteric coated polymer; and a method of using the controlled release oral solid formulation to treat Parkinson&#39;s disease or primary parkinsonism.

This application is a continuation of U.S. patent application Ser. No.16/360,936 filed on Mar. 21, 2019, which is a continuation of U.S.patent application Ser. No. 15/092,086 filed on Apr. 6, 2016, now U.S.Pat. No. 10,292,935, which is a continuation in part application of PCTApplication No. PCT/US2014/059554 filed Oct. 7, 2014, which claims thebenefit of U.S. Ser. No. 61/887,762 filed on Oct. 7, 2013, the contentsof all of which are hereby incorporated by reference in their entiretiesinto the present application.

Throughout this application various publications are referenced. Thedisclosures of these publications in their entireties are herebyincorporated by reference into this application in order to more fullydescribe the state of the art to which this invention pertains.

FIELD OF THE INVENTION

The present invention relates to controlled release pharmaceuticalcompositions of levodopa (LD) and esters of levodopa or salts thereof,formulated with a muco-adhesive polymer and an enteric coating polymerand, optionally, with a rate-controlling polymer, to yield enhanced drugdelivery attributes. These formulations are useful for the treatment ofconditions such as neurological diseases associated with reduced orimpaired dopamine levels.

BACKGROUND OF THE INVENTION

Patients suffering from Parkinson's disease (PD) frequently have periodsin which their mobility becomes difficult, often resulting in aninability to move. Abnormally low levels of dopamine, a neurotransmitterthat affects mobility and control of the skeletal-muscular system, iscommonly believed to be the main cause of these motor symptoms in PDpatients. However, administration of dopamine is not effective to treatthe motor symptoms of Parkinson's disease because dopamine does notcross the blood-brain barrier. To resolve this problem, PD patients areadministered levodopa, the metabolic precursor of dopamine, but levodopais not without its issues.

Over time patients treated with LD exhibit symptoms of “wearing off,”where a single dose of levodopa no longer lasts as long as in the earlydays of levodopa therapy (usually 5-10 years after start of levodopatherapy). Such patients may develop motor fluctuations characterized byend-of-dose failure, peak dose dyskinesia, and akinesia. The advancedform of motor fluctuations (also commonly referred to as the ‘on-off’phenomenon) is characterized by unpredictable swings from mobility toimmobility. Although the causes of these motor fluctuations are notcompletely understood, advanced patients generally benefit fromtreatment regimens that produce steady plasma levels of LD, such asthrough intestinal infusion of LD as such delivery method may mimicnormally tonic endogenous dopamine. However, intestinal infusion of LDis restrictive, invasive and cumbersome. Oral delivery of LD ispreferred, but plasma concentration levels remain difficult to controlvia oral delivery.

Combinations of levodopa (LD) and a decarboxylase inhibitor (typicallycarbidopa (CD)) to treat Parkinson's disease (PD) are known in thepharmaceutical arts. Currently, several formulations containing acombination of LD and CD are commercially available, e.g., SINEMET®,SINEMET® CR, STALEVO®, PARCOPA®, and their corresponding genericproducts. In addition, a decarboxylase inhibitor approved for useoutside of the United States, is benserazide, which may be given incombination with levodopa.

Nonetheless, a need remains for an oral LD formulation that providessteadier plasma concentrations of LD with minimal ‘peak-to-trough’fluctuations during daily dosing and that yields a longerduration-of-effect than the commercially available oral dosage forms ofLD. In addition, it is desirable for an oral LD formulation to providetherapeutic blood levels of LD quickly, thereby providing a rapid “on”to a PD patient in need thereof.

SUMMARY OF THE INVENTION

The current invention provides controlled release/extended absorptionoral dosage forms comprising levodopa and/or ester of levodopa or saltsthereof for treatment of Parkinson's disease and dopamine deficiencydisorders. More specifically, in some embodiments, the dosage formcomprises two types of components, the first component is an immediaterelease levodopa and/or its ester or salts thereof, and the secondcomponent comprises a core containing levodopa and/or ester of levodopaor salts thereof coated or layered with a muco-adhesive polymer andexternally coated with an enteric coating polymer. The dosage forms mayalso contain a rate controlling material that will contribute to thecontrolled release of the levodopa and/or ester of levodopa or saltsthereof. The rate controlling material may be a rate-controlling polymerapplied as an undercoating the muco-adhesive polymer coating or layer.The second component is essential to provide extended absorption,thereby providing prolonged and steady therapeutic coverage. The dosageform may comprise additionally a decarboxylase inhibitor, such ascarbidopa.

BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 shows the schematic configuration of the enteric-coated,muco-adhesive controlled release multi-particulates of this invention.

FIG. 2 is a line graph showing the in vitro dissolution profiles ofIPX203 multi-particulate formulations IPX203-C0004, IPX203-C0005 andIPX203-C0006.

FIG. 3 shows the plasma profile for IPX203 multi-particulateformulations IPX203-C0004, IPX203-C0005 and IPX203-C0006 in comparisonwith Sinemet® CR.

FIG. 4 are line graphs showing in vitro release profiles of testregimens A-D for IPX203-B13-01.

FIG. 5 is a line graph showing in vivo levodopa plasma profiles ofIPX203 formulations that provide plasma profiles with levodopa levelsmaintained at or greater than ½ Cmax longer than about 6 hours underfasted conditions.

FIG. 6 shows in vitro release profiles of IPX203-C0023, -C0024, -C0025and -C0026 formulations.

FIG. 7 shows in vivo levodopa plasma profiles for the formulationstested in IPX203-B14-01 PK study under fasted conditions.

DETAILED DESCRIPTION OF THE INVENTION

Unless defined otherwise, all technical and scientific terms used hereinhave the same meaning as commonly understood by one of ordinary skill inthe art to which the present disclosure belongs. As used herein thefollowing terms have the following meanings:

It must be noted that as used herein and in the appended claims, thesingular forms “a,” “an,” and “the” include plural referents unless thecontext clearly dictates otherwise. Thus, for example, reference to “aformulation” includes a plurality of compounds.

As used herein, the term “about” when used before a numericaldesignation, e.g., temperature, time, amount, concentration, and suchother, including a range, indicates approximations which may vary by (+)or (−) 10%, 5% or 1%.

Compositions of the Invention

The invention provides controlled release oral solid formulations oflevodopa and/or an ester of levodopa or a salt thereof providing arelatively steady levodopa plasma or serum concentration profile over aprolonged period of time and enhancing absorption of the active agentsin the gastrointestinal tract of a subject. Without being limited by anyone theory, it is believed that the polymer layers of the controlledrelease components of the present invention operate as follows. Theouter enteric coat delays release of the active agents until the dosageform has passed through the stomach and into the small intestine. In thesmall intestine, the muco-adhesive polymer facilitates adhesion to theintestinal mucosa, delaying passage of the dosage form through theintestine. It is desirable to retain the dosage form within the smallintestine where levodopa is absorbed most efficiently. In preferredembodiments, the rate controlling material further slows the release ofactive agent from the dosage form, thereby further extending the releaseand absorption of levodopa. Preferred formulations include an immediaterelease component to provide fast levodopa release and absorption, whichis important for PD patients in need of a fast “on.” As a result,formulations of the present invention can provide levodopa plasma levelsthat rise quickly and extend for a prolonged period of time.

Decarboxylase inhibitors such as carbidopa are often provided withlevodopa formulations in order to inhibit decarboxylation of levodopa,thereby increasing the levodopa bioavailability. In the formulations ofthe present invention, a decarboxylase inhibitor may be included in boththe immediate release component and the controlled release component.Preferably, the decarboxylase inhibitor is carbidopa and is includedonly in the immediate release component.

In one embodiment of the invention, the controlled release oral solidformulation contains (1) a controlled release component comprising alevodopa and/or an ester of levodopa or salts thereof and (2) animmediate release component comprising levodopa and/or an ester oflevodopa or salts thereof and a decarboxylase inhibitor. The immediaterelease component may be formulated as a powder, coating, mini-tablet,bead, pellet, granule or combination thereof. The controlled releasecomponent comprises a core containing levodopa and/or an ester oflevodopa or salts thereof coated with a layer of a muco-adhesive polymerand further coated with an outer layer of an enteric coating polymer.Preferably, the drug-containing core comprises a rate controllingmaterial, which may be mixed with the drug to form a controlled releasematrix core, coated onto the drug containing core to form an undercoatbelow the muco-adhesive polymer coating or layer, incorporated into themuco-adhesive polymer coating or layer, or a combination thereof. Insome embodiments, the immediate release component is in the form of apowder, pellet, bead or granule that is separate from the controlledrelease component. The immediate release component may also be appliedas an immediate release coating or layer onto the controlled releasecomponent.

In another embodiment of the invention, the controlled release oralsolid formulation contains (1) a controlled release component comprisinga levodopa and/or an ester of levodopa or salts thereof, and (2) adecarboxylase inhibitor component. The decarboxylase inhibitor componentmay be formulated as a powder, coating, mini-tablet, bead, pellet,granule or combination thereof. In this embodiment, the controlledrelease component comprises a drug-containing core with a layer orcoating of a muco-adhesive polymer and further coated with an outerlayer of an enteric coating polymer. Preferably, the drug-containingcore comprises a rate controlling material, which may be mixed with thedrug to form a controlled release matrix core, coated onto the drugcontaining core to form an undercoat below the muco-adhesive polymercoating or layer, incorporated into the muco-adhesive polymer coating orlayer, or a combination thereof. Preferably, the decarboxylase inhibitoris carbidopa.

The controlled release component may comprise drug-containing corescontaining both levodopa and/or an ester of levodopa or a salt thereofand a decarboxylase inhibitor, or the levodopa and/or ester of levodopaor salt thereof may be in separate controlled release components fromthat containing the decarboxylase inhibitor. Similarly, the immediaterelease component may also comprise a combination of levodopa and/or anester of levodopa or a salt thereof and a decarboxylase inhibitor. Thelevodopa and/or ester of levodopa or salt thereof may also be in aseparate immediate release component from the decarboxylase inhibitor.In one embodiment of the invention, the controlled release componentcomprises a levodopa-containing core free of a decarboxylase inhibitorsuch as carbidopa. Preferably, the formulation further comprises animmediate release component comprising levodopa and/or an ester oflevodopa or a salt thereof and a decarboxylase inhibitor.

In a preferred embodiment of the invention, the controlled release oralsolid formulation contains (1) a controlled release component comprisinglevodopa and/or ester of levodopa or salt thereof and (2) an immediaterelease component comprising levodopa and/or ester of levodopa or saltthereof and carbidopa. In this embodiment, the controlled releasecomponent comprises a drug-containing core coated with a first layer ofa rate controlling polymer, a second layer of muco-adhesive polymer andfurther coated with an outer third layer of an enteric coating polymer(see, e.g., FIG. 1).

In accordance with the practice of the invention, the formulations ofthe invention may be obtained by a granulation and/or coating processes,including, but not limited to, wet-granulation, fluid bedgranulation/coating or extrusion/spheronization, as is well-known in thepharmaceutical arts. The controlled release components and/or theimmediate release components may further contain conventionalpharmaceutically acceptable excipients such as lubricants, fillers,binders, disintegrants, glidants, surfactants (sometimes referred to aswetting agents), pH adjusting agents, antioxidants or mixtures of theforegoing.

In an embodiment of the invention, the controlled release and/orimmediate release components are multiparticulates that areencapsulated. The multiparticulates may be in a form that can besprinkled directly onto food or liquids for easy ingestion.

The active agents, such as decarboxylase inhibitor, levodopa and/orlevodopa ethyl ester, may be combined and dispersed throughout thedrug-containing core. In another embodiment, the active agents may bepresent in the center of the drug-containing core or layered/coated onan inert core such as a sugar sphere, microcrystalline cellulose sphere,glass sphere, plastic sphere or combination thereof.

In an embodiment of the invention, the controlled release oral solidformulation of levodopa or ester of levodopa or salts thereof maycomprise two or more controlled release components that release levodopaor ester of levodopa or salts thereof at different rates.

In this embodiment, the controlled release oral solid formulation oflevodopa or ester of levodopa or salts thereof may comprise at least twocontrolled release components differing in type, number, thicknessand/or composition of coating with a rate-controlling polymer, amuco-adhesive polymer and an enteric-coating polymer.

Examples of levodopa include but are not limited to levodopa, L-DOPA,L-3,4-dihydroxyphenylalanine, and(S)-2-amino-3-(3,4-dihydroxyphenyl)propanoic acid.

An example of a decarboxylase inhibitor includes, but is not limited to,carbidopa. Additional decarboxylase inhibitors include alpha methyldopa,benserazide (Ro4-4602), and alpha-difluoromethyl-DOPA (DFMD) or saltsthereof. In a preferred embodiment, the decarboxylase inhibitor iscarbidopa.

An example of an ester of levodopa is a levodopa ethyl ester (LDEE;ethyl (2S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate; CAS Number:37178-37-3) and having the structure:

(levodopa ethyl ester; CAS Number 37178-37-3).

Additional examples of esters of levodopa include, but are not limitedto:

levodopa butyl ester (butyl(2S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate; CAS Number: 39638-52-3)having the structure:

levodopa propyl ester; levodopa propyl ester (propyl(2S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate; CAS Number: 39638-51-2)having the structure:

and levodopa methyl ester (methyl(2S)-2-amino-3-(3,4-dihydroxyphenyl)propanoate; CAS Number: 7101-51-1),having the structure:

The ester of levodopa may be a salt, including, for example, a hydratedsalt. The salt of levodopa ester may comprise an organic or inorganicacid addition salt such as an octonoate salt, myristate salt, succinatesalt, succinate dihydrate salt, fumarate salt, fumarate dihydrate salt,mesylate salt, tartrate salt, sulfate salt, alkyl sulfate salt, maleatesalt, citrate salt, phosphate salt, acetate salt, lactate salt, nitratesalt, hydrochloride salt, hydrobromide salt, hydroidioide salt orcombination thereof.

For example, the succinate salt of an ester of levodopa or the succinatedihydrate salt may be a levodopa ethyl ester succinate (LDEE-S) orlevodopa ethyl ester succinate dihydrate (LDEE-S-dihydrate orLDEE-S(d)).

As used herein, “levodopa equivalence” or “LD equivalence” means thatamount of levodopa ester or salts thereof that contain equivalentamounts of levodopa, based on weight equivalence. For example, based onthe molecular weights, 306 mg of levodopa ethyl estersuccinate-dihydrate (LDEE-S-dihydrate) is equivalent to 228 mg oflevodopa ethyl ester (LDEE) and to 200 mg levodopa (LD).

Muco-adhesive polymers may be homogenous, i.e., a single type ofpolymer, or may comprise multiple types of muco-adhesive polymers.Muco-adhesive polymers may possess certain characteristics such as beinghydrophilic, hydrophobic, cationic, anionic and/or biocompatible andinclude multiple hydrogen bonding groups, hydrophobic surfaces,positively charged groups and/or negatively charged groups for adhesionto a mucosal surface so that the presence of active agent, such aslevodopa, can be prolonged at the site of absorption and increasebioavailability. Further, the muco-adhesive polymer may be natural,synthetic or from a biological source. Further still, the muco-adhesivepolymer may be composed of a single polymer or a combination of two ormore different polymers. In one embodiment, the polymers may range insize from 10,000 daltons to 1,000,000 daltons and more preferably 20,000daltons to 200,000 daltons.

An example of a muco-adhesive polymer includes, but is not limited to, abasic methacrylate copolymer, such as an amino methacrylate copolymer. Apreferred example of a methacrylate copolymer is a basic butylatedmethacrylate copolymer, an amino methacrylate copolymer, or aminoalkylmethacrylate copolymer, such as Eudragit® E 100

(poly(butyl methacrylate-co-(2-dimethylaminoethyl)methacrylate-co-methyl methacrylate) 1:2:1; CAS number: 24938-16-7;Evonik Industries). EUDRAGIT® E 100 is a cationic copolymer based ondimethylaminoethyl methacrylate, butyl methacrylate, and methylmethacrylate with a ratio of 2:1:1. The monomers are randomlydistributed along the copolymer chain. In a preferred embodiment, theaverage molar weight of EUDRAGIT® E 100 is approximately 150,000 g/mol.

Other examples of muco-adhesive polymer include, but are not limited to,a glyceride, steroidal detergent, polycarbophil (CAS Number 9003-97-8;Noveon® AA-1; Lubrizol Corp.), carbomer, cellulosics, chitosan

(CAS Number: 9012-76-4; Chitopharm® S with molecular weight range of50,000 to 1,000,000 daltons), diethylaminodextran,diethylaminoethyldextran, polygalactosamine, polylysine, polyomithine,prolamine, polyimine, hyaluronic acid, sodium alginate,hydroxypropylcellulose (HPC), hydroxypropylmethylcellulose (HPMC),sodium carboxymethylcellulose (sodium CMC) and alginate (CAS Number:9005-32-7) or combination thereof. Alginate is a homopolymer orheteropolymer composed of P-D-mannuronate (M) monomers, a-L-guluronate(G) monomers, or mixture of P-D-mannuronate and a-L-guluronate monomers

linked through (1→4) or (1,4)-glycosidic bonds. The (1,4)-glycosidiclinkages present in alginates are: β-D-mannuronate-(1,4)-β-D-mannuronate(MM), β-D-mannuronate-(1,4)-α-L-guluronate (MG),α-L-guluronate-(1,4)-β-D-mannuronate (GM) anda-L-guluronate-(1,4)-α-L-guluronate (GG), as can be seen below:

An alginate may be in the form of a polyanion or in the form of an acid,such as alginic acid. Further, alginate may be in the form of a salt ofalginic acid, such as sodium alginate, potassium alginate, ammoniumalginate, triethanolamine alginate, magnesium alginate or calciumalginate. Alternatively, alginate may be in the form of an ester ofalginic acid such as propylene glycol alginate.

The muco-adhesive polymer may constitute about 1-75% of the mass of thecontrolled release component, preferably about 2-70% of the mass of thecontrolled release component, most preferably about 3-50% of the mass ofthe controlled release component. Preferably, the muco-adhesive polymeris Eudragit® E 100. The muco-adhesive polymer percentages of mass statedabove are based on a multiparticulate with a bead size between 0.8 to1.2 mm. If the bead size is larger or smaller than 0.8 to 1.2 mm, theskilled artisan will understand that the mass percentage described aboveshould be adjusted accordingly.

Alternatively, the muco-adhesive polymer is a material capable offorming a positive ionic charge at the pHs present in the humangastro-intestinal tract. It is believed that the positive charge mayallow the muco-adhesive material to interact with the negative charge ofthe intestinal walls and thereby slow or delay the gastrointestinaltransit time of the controlled release component.

Enteric coating polymers are known in the art. In general, entericcoating polymers are designed to prevent drug release from an oral soliddosage form in the low pH environment of the stomach, thereby delayingdrug release until the dosage form reaches the small intestine. As such,the controlled release components of the invention have an in vitrorelease profile with minimal release of the active agent at pH 1.0. Inthe controlled release formulations of the invention, it is believed theouter enteric coating polymer layer provides an additional advantage inpreventing agglomeration of the controlled release components. That is,the enteric coat polymer layer prevents the controlled releasemuco-adhesive beads from sticking together in the low pH environment ofthe stomach.

The preferred enteric polymers are shellac (esters of aleurtic acid),zein, cellulose acetate phthalate (CAP), poly(methacrylic acid-co-methylmethacrylate), poly(methacrylic acid-co-ethyl methacrylate), celluloseacetate trimellitate (CAT), poly(vinyl acetate phthalate) (PVAP),hydroxypropyl methylcellulose phthalate (HPMCP) and hydroxypropylmethylcellulose acetate succinates. The preferred enteric polymersrelease at a pH of greater than or equal to pH 5.5. Examples includeEudragit® L 100 or Eudragit® L 100-55. The enteric coating polymers mayconstitute about 1-40% of the mass of the controlled release component,preferably about 1.5-30%, most preferably about 1.5-25%. Theenteric-coated polymer percentages stated above are based on amultiparticulate bead size between 0.8-1.2 mm. If the bead size issmaller or larger, the skilled artisan will understand that the masspercentage described above should be adjusted accordingly.

The enteric coating should be designed to dissolve at a pH greater than5.0, preferably at a pH of 6.0 or higher and most preferably a pH of 6.5or higher.

The enteric coating polymer may comprise a methacrylic acid copolymer ormultiple types of methacrylic acid copolymers. The methacrylic copolymermay comprise any of Eudragit® L 30 D-55

(poly(methacrylic acid-co-ethyl acrylate) 1:1; CAS Number 25212-88-8;Evonik Industries), Eudragit® L 100-55

(poly(methacrylic acid-co-ethyl acrylate) 1:1; CAS Number 25212-88-8;Evonik Industries), Eudragit® L 100

(poly(methacrylic acid-co-methyl methacrylate) 1:1; CAS Number25086-15-1; Evonik 15 Industries), Eudragit® L 12.5

(poly(methacrylic acid-co-methyl methacrylate) 1:1; CAS Number25086-15-1; Evonik Industries); Eudragit® S 100

(poly(methacrylic acid-co-methyl methacrylate) 1:2; CAS Number25086-15-1; Evonik Industries), Eudragit® S 12.5

(poly(methacrylic acid-co-methyl methacrylate) 1:2; CAS Number25086-15-1; Evonik Industries), and Eudragit® FS 30 D

(poly(methyl acrylate-co-methyl methacrylate-co-methacrylic acid) 7:3:1;CAS Number 26936-24-3; Evonik Industries) or a combination thereof.

In a preferred embodiment of present invention, the controlled releasecomponent comprises a further rate-controlling polymer coat over thedrug-containing core, undercoating the muco-adhesive polymer. Examplesof rate-controlling polymers useful in the present invention include,but are not limited to, ethylcellulose, cellulose acetate, Eudragit® E,Eudragit® RS (ammonio methacrylate copolymer), Eudragit® RL (ammoniomethacrylate copolymer), and Eudragit® NE (a neutral copolymer ofpolymethacrylic acid esters), or mixtures thereof. Preferably, therate-controlling polymers are not soluble in water at neutral pH.Additional rate-controlling polymers that may be used are described inU.S. Pat. No. 5,002,776 which is incorporated herein by reference.Preferably, the rate-controlling polymer is cellulose acetate orethylcellulose. The rate-controlling polymer can also include a poreforming agent or a flux enhancer to adjust the release rate. Preferably,the pore forming agent or flux enhancer is a water soluble material suchas a salt, i.e., NaCl, KCl, a sugar, i.e., lactose, sucrose, mannitol,povidone, copovidone, polyethylene glycol, hydroxypropyl cellulose,hydroxypropyl methylcelluose or combinations thereof. If the poreforming agent or flux enhancer is a water soluble polymer, it shouldhave a low molecular weight such as below 100,000, preferably below50,000 and/or should rapidly dissolve in water, i.e., 2 wt % of thewater soluble polymer should dissolve in 100 ml of water within 15minutes or less, preferably 10 minutes or less, and most preferably 5minutes or less at 25° C.

The controlled release component may also comprise a hydrophobic ratecontrolling material in addition to or in place of the rate-controllingpolymer. Examples of hydrophobic materials that can be used includebeeswax, white wax, emulsifying wax, hydrogenated vegetable oil,hydrogenated castor oil, microcrystalline wax, cetyl alcohol, stearylalcohol, free wax acids such as stearic acid, esters of wax acids,propylene glycol monostearate, glycerol monostearate, carnauba wax, palmwax, candelilla wax, lignite wax, ozokerite, ceresin wax, lardaceine,China wax and mixtures thereof. Other possible rate-controllingexcipients useful in the present invention include saturatedhydrocarbons having from 25 to 31 carbon atoms, saturated alcoholshaving from 25 to 31 carbon atoms, saturated monocarboxylic acids havingfrom 25 to 31 carbon atoms, esters obtained from said alcohols, andmonocarboxylic acids which are described in U.S. Pat. No. 6,923,984,incorporated herein by reference.

In an alternate embodiment, the controlled release component comprises amatrix core comprising a mixture of a controlled release material, whichmay be the afore-described rate-controlling polymer and/or hydrophobicmaterial and the drug, i.e., levodopa and/or ester of levodopa or saltsthereof and/or decarboxylase inhibitor. The matrix core may furthercomprise one or more pharmaceutically acceptable excipients such aslubricants, fillers, binders, disintegrants, glidants, surfactants(sometimes referred to as wetting agents), pH adjusting agents,antioxidants or mixtures of the foregoing. In this embodiment, thematrix core may be further coated with a rate controlling coating orpolymer before being coated with the muco-adhesive coating and the outerenteric coating.

In another alternate embodiment, the controlled release component mayincorporate a controlled release material, which may be theafore-described rate-controlling polymer and/or hydrophobic material,into the muco-adhesive coating.

The controlled release material may constitute about 1-35% of the massof the controlled release component, preferably about 2-30% and mostpreferably about 3-25%.

The muco-adhesive coating or layer and enteric coatings employed in thepresent invention may further comprise one or more pharmaceuticallyacceptable excipients such as plasticizers, lubricants, fillers,binders, disintegrants, glidants, surfactants (sometimes referred to aswetting agents), pH adjusting agents, antioxidants, or mixtures of theforegoing.

Some commonly known plasticizers include adipate, azelate, enzoate,citrate, stearate, isoebucate, sebacate, triethyl citrate, tri-n-butylcitrate, acetyl tri-n-butyl citrate, citric acid esters, and thosedescribed in the Encyclopedia of Polymer Science and Technology, Vol. 10(1969), published by John Wiley & Sons. The preferred plasticizers aretriacetin, acetylated monoglyceride, grape seed oil, olive oil, sesameoil, acetyltributylcitrate, acetyltriethylcitrate, glycerin sorbitol,diethyloxalate, diethylmalate, diethylfumarate, dibutylsuccinate,diethylmalonate, dioctylphthalate, dibutylsebacate, triethylcitrate,tributylcitrate, glyceroltributyrate and combinations thereof. Dependingon the particular plasticizer, amounts from about 0% to about 25%, andpreferably about 2% to about 15%, of the plasticizer can be used basedupon the total weight of the controlled release, muco-adhesive and/orenteric coating.

Lubricants useful in pharmaceutical formulations are known in the art.Examples of a suitable lubricant include, but are not limited to,stearic acid, lauric acid, myristic acid, palmitic acid, fatty acid,magnesium stearate, calcium stearate, zinc stearate, sodium stearate,Stear-O-Wet®, sodium stearyl fumarate, salt of a fatty acid, metallicsalt of fatty acid, glyceryl monostearate, glyceryl tribehenate,glyceryl dibehenate, Compritol® 888 ATO, glyceride ester, sorbitanmonostearate, sucrose monopalmitate, sugar ester, fatty acid ester,talc, hydrated magnesium silicate, PEG 4000, boric acid, Carbowax (PEG)4000/6000, sodium oleate, sodium benzoate, sodium acetate, sodium laurylsulfate, magnesium lauryl sulfate, Sterotex, wax, or mixture thereof.

Examples of fillers that may be employed in the composition of thepresent invention include sugars, such as lactose, sucrose, mannitiol,dibasic calcium phosphate, microcrystalline cellulose, calciumcarbonate, magnesium carbonate, calcium sulfate, powdered cellulose,silicified microcrystalline cellulose, magnesium carbonate, magnesiumoxide, starch, and mixtures thereof. The filler may constitute about1-50% of the mass of the controlled release component, preferably about2-45% and most preferably about 5-40%.

Examples of binders that may be employed in the compositions of thepresent invention include acacia, povidone, hypromellose, hydroxypropylcellulose, hydroxyethyl cellulose, polyethylene oxide,polymethacrylates, methyl cellulose, ethyl cellulose, pregelatinizedstarch, gelatin, tragacanth, zein, or mixtures thereof. Preferably, thebinder is selected from povidone, hypromellose, hydroxypropyl cellulose,hydroxyethyl cellulose, polymethacrylates, methyl cellulose, gelatin andethyl cellulose, or mixtures thereof. Especially preferred bindersinclude water soluble binders such as povidone, hypromellose,hydroxypropyl cellulose, gelatin and mixtures thereof. The binder mayconstitute about 0.1-15% of the mass of the controlled releasecomponent, preferably about 0.2-10% and most preferably about 0.5-5%.

Examples of disintegrants that may be employed in the compositions ofthe present invention include croscarmellose sodium, starch,crospovidone, sodium starch glycolate, alginic acid, calciumcarboxymethylcellulose, sodium carboxymethylcellulose, calciumcarboxymethylcellulose, powdered cellulose, chitosan, guar gum,magnesium aluminium silicate, methylcellulose, sodium alginate, andmixtures thereof.

Examples of glidants that may be employed in the compositions of thepresent invention include colloidal silicon dioxide, cornstarch, talc ormixtures thereof.

One or more surfactants may also be employed in the compositions of thepresent invention. The surfactant may be a non-ionic surfactant or anionic surfactant. Examples of non-ionic surfactants includepolyethoxylated castor oil, a polyoxyethylene alkyl ester, apolyglycolyzed glyceride, a sorbitan fatty acid ester, a glycerin fattyacid ester, a fatty acid polyglyceride, a fatty acid alcohol polyglycolether, acetylene glycol, acetylene alcohol, an oxyalkylene blockpolymer, a polyoxyethylene alkyl ether, a polyoxyethylene alkylarylether, a polyoxyethylene styrylaryl ether, a polyoxyethylene glycolalkyl ether, a polyoxyethylene fatty acid ester, a polyoxyethylenesorbitan fatty acid ester, a polyoxyethylene glycerin fatty acid ester,a polyoxyethylene hydrogenated castor oil, a polyoxypropylene fatty acidester, or a mixture of the foregoing. A further listing of possiblenon-ionic surfactants can be found on pages 1243-1249 of Martindale, TheExtra Pharmacopoeia 29^(th) ed. which is incorporated herein byreference.

In certain embodiments, the non-ionic surfactants may comprise fattyalcohol acid or amide ethoxylates, monoglyceride ethoxylates, sorbitanester ethoxylates alkyl polyglycosides, mixtures thereof, and the like.Certain non-ionic surfactants include polyoxyethylene derivatives ofpolyol esters, such as Polysorbate 20 (TWEEN 20®), Polysorbate 40 (TWEEN40®) Polysorbate 60 (TWEEN 60®), and Polysorbate 80 (TWEEN 80®).

In certain embodiments, the non-ionic surfactant may also comprised-alpha tocopheryl polyethylene glycol 1000 succinate (TPGS),nonoxinols, poloxamers, sorbitan monolaurate, sorbitan monooleate,sorbitan monopalmitate, sorbitan monostearate, sorbitan sesquioleate,sorbitan trioleate, tyloxapol, and mixtures of the foregoing.

Any variety of ionic surfactants may also be incorporated into thecompositions of the present invention. Suitable ionic surfactantsinclude, but are not limited to, carboxylates such as soaps, acyllactylates, acyl amides of amino acids, esters of sulfuric acid such asalkyl sulfates and ethoxylated alkyl sulfates, sulfonates such as alkylbenzene sulfonates, acyl isethionates, acyl taurates andsulfosuccinates, phosphates, quaternary ammonium salts, and ethoxylatedamines. An example of a preferred ionic surfactant is sodium laurylsulfate.

Examples of pH adjusting agents that may be employed in the compositionsof the present invention include pharmaceutically acceptable acids orbases which may be present to adjust the pH of intermediate compositionsleading up to the final compositions and to adjust the pH of the drugenvironment of final compositions to a desired or optimum pH range.Representative examples of pharmaceutically acceptable acids that may beused include, but are not limited to, acetic acid, citric acid, fumaricacid, hydrochloric acid, malic acid, nitric acid, phosphoric acid,propionic acid, sulfuric acid, tartaric acid, and mixtures thereof.Representative examples of pharmaceutically acceptable bases that may beused include but are not limited to ammonia, ammonium carbonate,diethanolamine, potassium hydroxide, sodium bicarbonate, sodiumcarbonate, sodium hydroxide, trolamine, and mixtures thereof. In certainembodiments the pH adjusting agent is an acid, preferably an organicacid and will constitute about 0.5-20% of the mass of the controlledrelease component, preferably about 0.75-15% and most preferably about1-10%. Alternatively, the pH adjusting agent is an acid, preferably anorganic acid and will be present in the controlled release component ina molar ratio of acid to levodopa of about 1:4 to about 4:1, preferablyabout 1:3 to about 3:1 and most preferably about 1:2 to about 2:1.

Examples of antioxidants that may be employed in the compositions of thepresent invention include ascorbic acid, ascorbyl palmitate, butylatedhydroxyanisole, butylated hydroxytoluene, hypophosphorous acid,monothioglycerol, potassium metabisulfate, propyl gallate, sodiumbisulfite, sodium formaldehyde sulfoxylate, sodium metabisulfate, sodiumsulfate, sodium thiosulfate, sodium dioxide, tocopherol, and mixturesthereof.

In an embodiment of the invention, the carbidopa and the levodopa orlevodopa equivalence are present in the formulation of the invention ina weight ratio of about 1:1 to about 1:10, preferably about 1:4.

For example, useful amounts of levodopa or levodopa equivalence andcarbidopa include: (a) 200 mg:31.25 mg; (b) 200 mg:50 mg; (c) 255.6mg:50 mg; (d) 360 mg:50 mg; (e) 95 mg:23.75 mg; (f) 145 mg:36.25 mg; (g)195 mg:48.75 mg; (h) 245 mg:61.25 mg; or (i) 390 mg:97.5 mg; with eachvalue capable of varying by ±10%. Further examples include amounts oflevodopa:carbidopa or levodopa equivalence:carbidopa as follows: (a) 95mg:23.75 mg; (b) 145 mg:36.25 mg; (c) 195 mg:48.75 mg; or (d) 245mg:61.25 mg; with each value capable of varying by ±10%.

In an embodiment of the invention, the immediate release component maycomprise less levodopa or levodopa equivalence dosage than thecontrolled release component. For example, the ratio of levodopa orlevodopa equivalence in the immediate release component to that in thecontrolled release component can be in the range of 0.15 to 0.65. Forexample, a ratio in weight of levodopa equivalence in the controlledrelease component:immediate release component is at least about 2:1,most preferably 3:1. Preferably the amount of levodopa or ester oflevodopa or salts thereof in the immediate release component shouldprovide a therapeutic dose of levodopa or ester of levodopa or saltsthereof within one hour or less after administration of the finalcomposition of the present invention, preferably within 45 minutes orless after administration and most preferably about 30 minutes or lessafter administration.

In one embodiment of the invention, the controlled release component isa plurality of beads, pellets or granules having a size that passesthrough 12, 14, or 16 mesh but may be retained on 18, 24 or 25 meshscreens. Further, the beads, pellets, or granules may have a size thatpasses through 14 mesh but may be retained on 18 or 24 mesh screens.

The controlled release component will have an in vitro dissolutionprofile showing minimal release of the levodopa and/or ester of thelevodopa or a salt thereof at pH 1.0 and extended release of the esterof levodopa or a salt thereof near neutral pH, for example at or near pH7. For example, minimal release may entail less than 20% release oflevodopa, preferably less than 10%, most preferably less than 5% usingUnited States Pharmacopeia (USP) I dissolution method at agitation speedof 75 rpm in Simulated Gastric Fluid (pH 1.0, without enzyme) for 2 hrs.Further, extended release may involve release at over at least four andup to an additional 8 hours at or near pH 7, upon changing to SimulatedIntestinal Fluid (pH 7.0, without enzyme) after first 2 hrs in SimulatedGastric Fluid (pH 1.0, without enzyme) using USP I dissolution method atagitation speed of 75 rpm. Further still, as used here, at or near pH 7includes a pH at or about pH 6.5, 6.6, 6.7, 6.8 6.9, 7.1, 7.2, 7.3, 7.4,7.5 or 7.6.

In certain embodiments, the composition should comprise an immediaterelease component and at least one controlled release component whereinfollowing administration to a human patient the immediate releasecomponent should provide a therapeutic dose of levodopa or ester oflevodopa or salts thereof within one hour or less after administrationof the final composition of the present invention, preferably within 45minutes or less after administration and most preferably about 30minutes or less after administration. After the thereapeutic plasmalevel of levodopa is obtained by the immediate release component, thecontrolled release component(s) should provide and/or maintain atherapeutic plasma level of levodopa or ester of levodopa or salts overa period of thereof at least 4-12 hours, preferably 6-12 hours and mostpreferably 5, 6, 7, 8, 9, 10, 11 or 12 hours after administration. Toobtain this therapeutic level, the controlled release component(s)should exhibit the following in vitro release profiles when tested usinga USP Type I or II apparatus, at 37° C., with a rotational speed of 75rpms and 900 ml of an aqueous media with a pH between 6.8-7.4 andpreferably at a pH of 7:

Levododopa Released Time Preferred More Preferred Most Preferred 2 0-60% 10-55% 15-50% 4  25-90% 30-85% 35-80% 6 35-100% 40-100%  50-100% 

The amount of levodopa released in the above table is based on the totalamount of levodopa or ester of levodopa or salt thereof present in thecontrolled release component. In certain embodiments the controlledrelease component(s) should also release less than 25% of the levodopaor ester of levodopa or salt, preferably less than 20 and mostpreferably less than 15% when tested using a USP Type I or II apparatus,at 37° C., with a rotational speed of 75 rpms and 900 ml of an aqueousmedia with a pH of 1.

In a further embodiment, the present invention comprises: a) animmediate release component as previously described; b) a controlledrelease component as previously described and c) an enteric coatedcomponent. The enteric coated component comprises a core comprisinglevodopa or ester of levodopa or salt thereof and/or a decarboxylaseinhibitor and at least one pharmaceutically acceptable excipient aspreviously described and an enteric coating. The enteric coatedcomponent will release 100% of the levodopa or ester of levodopa or saltthereof and/or a decarboxylase inhibitor within 90 minutes, preferably60 minutes and most preferably within 45 minutes when tested using a USPType I or II apparatus, at 37° C., with a rotational speed of 75 rpmsand 900 ml of an aqueous media with a pH between 6.8-7.4, preferably atpH 7. The enteric coated component will also release less than 25% ofthe levodopa or ester of levodopa or salt, preferably less than 20% andmost preferably less than 15% when tested using a USP Type I or IIapparatus, at 37° C., with a rotational speed of 75 rpms and 900 ml ofan aqueous media with a pH of 1.

The levodopa and/or ester of levodopa or a salt thereof released fromthe controlled release component may produce an in vivo levodopa plasmaprofile (e.g., mean in vivo levodopa plasma profile) comprising a peakoccurring not before about two hours after administration to a subjectand provides at least three hour duration for levodopa plasmaconcentration above 50% the maximum value of the peak concentration(Cmax). In another embodiment, in the plasma profile, the peak occursafter about one and a half hours after administration to the subject andexhibits at least a four-hour duration for levodopa plasma concentrationat or above 50% of Cmax. By way of example, the profile may be achievedunder fasting conditions.

When the formulation of the invention comprises an immediate releasecomponent and a controlled release component, the in vivo levodopaplasma profile following administration of an oral dosage form of theformulation to a subject may comprise time of administration of an oraldosage form; a levodopa plasma concentration corresponding to Cmaxoccurring within about 6 hours or 7 hours after administration of thedosage form; a mean time to reach 50% of Cmax within one hour ofadministration, more preferably within 30 minutes. The time to 50% ofCmax is less than one hour and 50% Cmax is maintained for at least 5.0hours. The time after administration of the dosage form when the maximumplasma concentration is reached (Tmax) is between 30 minutes and 7hours. Preferably, the LD plasma level is maintained at or above 50% ofCmax for at least 5.5 hours, more preferably, for at least 6.0 hours,even more preferably, for at least 6.5 hours, and most preferably for atleast 7.0 hours.

In one embodiment, the formulations of the invention may have a ratio ofsaid Cmax to the mass of levodopa or levodopa equivalence. Theconcentration may be measured in units of ng/mL, to the mass of levodopaor levodopa equivalence in the formulation, where said mass is measuredin mg, of between 2:1 and 6:1. The ratio may be between 2.5:1 and 5.5:1,preferably, greater than or equal to about 3:1.

The combination of immediate release components and controlled releasecomponents of the invention provide the near infusion-like profile asevident from the plateau in the LD plasma profile (see, e.g., FIG. 5).The LD Cmax itself is not clinically relevant. What is clinicallyrelevant is the time to reach a therapeutic level of LD (e.g., an LDlevel of 50% Cmax) and the time maintained at or above the therapeuticlevel (e.g., 50% Cmax). The short time to reach a therapeutic LD levelis associated with a faster “on” time for PD patients, whereas theprolonged period at or above therapeutic levels provides the desiredsteady “infusion-like” profile.

It is an advantage of the present invention to provide a sustainedlevodopa plasma concentration for a duration greater than 5 hrs and amore consistent duration with percent coefficient of variation (CV) ofmean duration of levodopa plasma concentrations >50% C_(max) of lessthan 35%, preferrably less than 30%.

The skilled artisan will appreciate that daily dosages having an amountof active agent sufficient or effective to treat diseases associatedwith reduced or impaired dopamine levels may generally contain fromabout 25 mg to about 6000 mg of levodopa or levodopa equivalence dose incombination with from about 5 mg to about 1500 mg of carbidopa.

Dosage forms may contain 25-750 mg of levodopa or levodopa equivalence.Further, dosage forms may contain carbidopa ranging from 25-300 mg. Forexample, the controlled release oral solid formulation of the inventionmay comprise from about 25 mg to about 1000 mg levodopa or levodopaequivalence. Further, the controlled release oral solid formulation ofthe invention may comprise from about 10 mg to about 300 mg carbidopa.Further still, the controlled release oral solid formulation of theinvention may comprise from about 10 mg to about 150 mg carbidopa.

By way of example, the total daily dose of levodopa from theformulations of the invention may be less than about 2500 mg. Forexample, the total daily levodopa dose may be between 800 mg to 2500 mg.In a further example, the total daily levodopa dose may be about 855 mg,1140 mg, 1170 mg, 1305 mg, 1755 mg, 2205 mg, or 2340 mg. In anotherembodiment, the total daily carbidopa dose may be about 292.5 mg.

The dosing frequency may vary, depending on the need of the subject. Forexample, the dosing frequency of the formulations of the invention maybe two, three, four or five times a day preferably two or three times aday. In another example, the dosing frequency may be a maximum of fivetimes a day.

Actual dosage levels of active ingredient in the compositions of thepresent invention may be varied so as to obtain an amount of activeingredient that is effective to obtain a desired therapeutic responsefor a particular composition. The formulations of the invention may beadministered as a single dose, or may comprise of a number of smallerdoses, such as two 100 mg dose of levodopa for a total 200 mg dose, tobe administered or consumed within a short period of time, such aswithin 15 minutes or less, preferably within 10 minutes or less and mostpreferably within 5 minutes of less. It is understood that the precisedosage and duration of treatment is a function of the disease beingtreated and may be determined using known practices. It is to be notedthat dosage values may also vary with the severity of the condition tobe alleviated. It is to be further understood that for any particularsubject, specific dosage regimens should be adjusted over time accordingto the individual need and the professional judgment of the personadministering or supervising the administration of the formulations ofthe invention, and that the concentration ranges set forth herein areexemplary only and are not intended to limit the scope or practice ofthe claimed compositions. Preferably the doses may be administered 2, 3or 4 times a day (per 24 hour time period), and most preferably 2 or 3times a day.

Optimally, after administration to a patient suffering from a conditionassociated with reduced or impaired dopamine levels, a pharmaceuticalformulation of the invention releases levodopa into the plasma of thepatient at a steady or near constant level without significant decreaseor fluctuation for an extended amount of time, thereby reducing motorfluctuations.

The invention also provides methods for treating a subject withParkinson's disease or primary Parkinsonism. The method comprisesadministering to the subject an effective amount of any of thecontrolled release oral solid formulations of the invention to treatParkinson's disease or primary Parkinsonism. In accordance with thepractice of this invention, the subject may be a human.

Preferred compositions of the present invention include:

A multiparticulate formulation comprising: a) a immediate releasecomponent of levodopa or ester of levodopa or salt thereof and/or adecarboxylase inhibitor and optionally at least one pharmaceuticallyacceptable excipient as previously described and b) a controlled releasecomponent comprising a plurality of controlled release beads, pellets orgranules wherein the beads, pellets or granules comprise a core oflevodopa or ester of levodopa or salt thereof and/or a decarboxylaseinhibitor and at least one pharmaceutically acceptable excipient aspreviously described, a muco-adhesive coating or layer applied to and/orsurrounding the core and an enteric coating surrounding themuco-adhesive coating or layer wherein the controlled release beadspellets or granules also comprises a controlled release material. Thecontrolled release material may be: i) mixed with the levodopa or esterof levodopa or salt thereof and/or a decarboxylase inhibitor and atleast one pharmaceutically acceptable excipient to form a controlledrelease matrix core, ii) applied as a coating or layer onto the corecomprising the levodopa or ester of levodopa or salt thereof and/or adecarboxylase inhibitor and at least one pharmaceutically acceptableexcipient; iii) incorporated or mixed into the muco-adhesive coating orlayer or iv) a combination of (i), (ii) and/or (iii).

An alternative preferred compositions includes:

a multiparticulate formulation comprising: a) a immediate releasecomponent of levodopa or ester of levodopa or salt thereof and/or adecarboxylase inhibitor and optionally at least one pharmaceuticallyacceptable excipient as previously described; b) a controlled releasecomponent comprising a plurality of controlled release beads, pellets orgranules wherein the beads, pellets or granules comprise a core oflevodopa or ester of levodopa or salt thereof and/or a decarboxylaseinhibitor and at least one pharmaceutically acceptable excipient aspreviously described, a muco-adhesive coating or layer applied to and/orsurrounding the core and an enteric coating surrounding themuco-adhesive coating or layer wherein the controlled release beadspellets or granules also comprises a controlled release material and c)an enteric coated component comprising a plurality of enteric coatedbeads pellets or granules comprising a core comprising levodopa or esterof levodopa or salt thereof and/or a decarboxylase inhibitor and atleast one pharmaceutically acceptable excipient as previously describedand an enteric coating surrounding the core. The controlled releasematerial employed in the controlled release component may be: i) mixedwith the levodopa or ester of levodopa or salt thereof and/or adecarboxylase inhibitor and at least one pharmaceutically acceptableexcipient to form a controlled release matrix core, ii) applied as acoating or layer onto the core of the levodopa or ester of levodopa orsalt thereof and/or a decarboxylase inhibitor and at least onepharmaceutically acceptable excipient; iii) incorporated or mixed intothe muco-adhesive coating or layer or iv) a combination of (i), (ii)and/or (iii). The enteric coated component will release 100% of thelevodopa or ester of levodopa or salt thereof and/or a decarboxylaseinhibitor within 90 minutes, preferably 60 minutes and most preferablywithin 45 minutes when tested using a USP Type I or II apparatus, at 37°C., with a rotational speed of 75 rpms and 900 ml of an aqueous mediawith a pH between 6.8-7.4.

The multiparticulate formulations may be in the form or a tablet orcapsule or may be sprinkled on to food such as applesauce foradministration to a patient suffering from Parkinson's disease.

EXAMPLES Example 1

I. Development of LDEE-S beads for IPX203-B-12-01

Development of Core LDEE-S Breads

Preparation of Core Beads

Required amounts of LDEE-S-Dihydrate, Microcrystlline Cellulose, Fumaricacid, Povidone K29-32, ethanol and Purified Water were dispensed. Thealcohol and the purified water were charged into a container and stirredusing a stir bar. Povidone was slowly added into the ethanol/water mixedsolvent. Mixing continued until the Povidone was completely dissolved,and the spray pump was calibrated to the target granulation spray rate.

LDEE-S-Dihydrate, Microcrystalline Cellulose, Fumaric acid, and Povidonewere charged into a high shear granulator and dry mixed for 1-5 minutesat impeller speed of 75 rpm and chopper speed of 1000 rpm. The Povidonesolution was sprayed into the granulation bowl and granulation continuedwith either ethanol or water as necessary. The granules were wet mixedfor 2 minutes, after the spraying was completed.

The wet granules were extruded using the extruder (MG 55 MultiGranulator) equipped with a 0.8 mm hole size screen at extruder speed of55 rpm. The extrudates were collected into double polyethylene linedbags. The collected extrudate was weighed and adjusted in the quantitiesranging from 170-210 g per load.

One load of the weighed extrudate was charge into a spheronizer equippedwith a 3 mm cross hatch disc. The extrudate was spheronized at aspheronisation speed of 1400 rpm for 1-10 mins. The spheronized beadswere discharge into double polyethylene bags. The remaining extrudatewere spheronized until all the double polyethylene-lined bags arecompleted.

The wet beads were dried in a fluid bed drier (Glatt GPCP-1) at an Inlettemperature of 35±10° C. until Loss on Drying was not more than 5.0%.The steps above were repeated until additional sub loads had beenprocessed.

The dried beads were passed through a mechanical sieve (Vibroscreen)equipped with a 24-MG mesh screen at the bottom, 18-MG mesh screen inthe middle, and 16-MG mesh screen at the top. The beads that remained on18-US mesh and 24-MG mesh screens were collected into doublepolyethylene lined bags.

Muco-Adhesive/Rate-Controlling Sub-Layer Coating

The batch yield was determined. The required amounts of AminoMethacrylate Copolymer (Eudragit® E 100) and Talc were calculated anddispensed. Purified Water, Acetone and Isopropyl Alcohol were dispensedinto a stainless steel container and stirred using stir bar. Whilestirring, Amino Methacrylate Copolymer (Eudragit® E 100) was slowlyadded into the vortex of the mixed solvent. Mixing continued until thecopolymer completely dissolved. While stirring, Talc was slowly disperseinto the vortex of the solution. Mixing continued until the material wascompletely dispersed. The suspension was continually stirred throughoutthe coating process.

The spray pump was calibrated to the target coating spray rate of theperistaltic pump using the suspension solution above. The core beadswere coated using Glatt GPCG 1 equipped with a Wurster insert at Inletair temperature of 35±10° C., Atomization air pressure of 1.0-2.0 barsand Wurster partition height of 15-30 mm. During coating, the inlet airtemperature, exhaust flap, and spray rate were adjusted to maintain theexhaust air temperature between 30±5° C.

After the target amount of coating solution was sprayed, the coatedbeads were dried at an inlet air temperature of 40±10° C. for 90minutes. The dried beads were passed through a mechanical sieve(Vibroscreen) equipped with a pan at the bottom, 14-MG mesh screen inthe middle, and 12-MG mesh screen at the top. The beads that remained inthe pan and 14-MG mesh screens were collected into double polyethylenelined bags.

Enteric Coating

The batch yield was determined. Based on the batch yield, the requiredamounts of Triethyl Citrate, Talc and an enteric copolymer, eitherMethacrylic Acid Copolymer, Type A, (Eudragit® L 100)/Methacrylic AcidCopolymer, Type B, (Eudragit® S) at ½ weight ratio for IPX203-C0006 orEudragit® L 100-55 for IPX203-C0004 and IPX203-C0005 were calculated anddispensed. Acetone and Isopropyl Alcohol for IPX203-C0006 or Acetone,Isopropyl Alcohol and purified water for IPX203-C0004 and IPX203-C0005were dispensed into a stainless steel container and stirred using stirbar. While stirring, the enteric copolymer and Triethyl Citrate wereadded slowly into the vortex of the mixed solvent. Mixing continueduntil the copolymer was completely dissolved.

While stirring, Talc was added slowly into the vortex of the solution.Mixing continued until the material was completely dispersed. Thesuspension was continually stirred throughout the coating process. Thespray pump was calibrated to the target coating spray rate of theperistaltic pump using the suspension solution.

Eudragit® E-coated beads were coated with the enteric composition usingGlatt GPCG 1 equipped with a Wurster insert at Inlet air temperature of35±10° C., Atomization air pressure of 1.0-2.0 bars and Wursterpartition height of 15-30 mm. During coating, the inlet air temperature,exhaust flap, and spray rate were adjusted to maintain the exhaust airtemperature between 30±5° C.

After the target amount of coating solution was sprayed, theenteric-coated beads were dried at an inlet air temperature of 40±10° C.for 120 minutes. The dried beads were passed through a mechanical sieve(Vibroscreen) equipped with a pan at the bottom, 14-MG mesh screen inthe middle, and 12-MG mesh screen at the top. The beads that remained inthe pan and 14-MG mesh screens were collected into double polyethylenelined bags.

Encapsulation

The batch yield was determined. Based on the batch yield, the requiredamounts of the enteric coated beads (also referred to herein as beadshaving an outer enteric coating polymer layer) and talc were calculatedand dispensed. The Enteric Coated Beads and Talc were placed in anappropriated sized plastic bag and were manually blended by shaking theplastic bag with the beads and Talc for 10 minutes. The blend wasencapsulated with 00 size gelatin capsules, using MG FlexalabEncapsulator at the target fill weight of 482 mg, 537 mg and 472 mg forIPX203-C0004, IPX203-C0005 and IPX203-C0006 respectively, so that thetarget LDEE dose/2 capsules was 228 mg, equivalent to LD dose of 200 mg.

Rationale for Components and Coatings in Formulation

The core bead formulation was developed utilizing microcrystallinecellulose (MCC) as filler since the wetted MCC has the desiredrheological properties, cohesiveness, and plasticity to yield strongbeads. An MCC level at 30% was selected and it was found to providebeads with acceptable sphericity and support a robust manufacturingprocess. Because LDEE-S is more stable in acidic environment, in orderto reduce the LDEE degradation inside the beads during the long releaseduration, a 5% fumaric acid is added in the formulation to lower themicroenvironment pH. An extra binder povidone at 1% level is also addedto the formulation with the intent to provide a more robust extrusionprocess. The dissolution profile of the core beads is fast, with thecomplete release within 30 min, as measured in a USP Apparatus 1 withbasket speed of 75 rpm in pH 7 phosphate buffer.

To control the release of LDEE-S, the core LDEE-S beads are coated withdifferent release polymers. Eudragit® E 100 is swellable and permeableabove pH 5. It is used as an inner coating to slowly release drug atintestinal pH. As such, the use of Eudragit® E 100 coating results in acontrolled release of LDEE-S. Furthermore, to protect the Eudragit® Elayer as well as to direct the release of LDEE-S to the more alkalineregion (i.e., intestinal region and not the stomach region), an entericcoating is applied as an outer coat.

Development of Eudragit® E 100 Coated LDEE-S Beads

Prototype formulations with different Eudragit® E 100 coating contentwere developed and evaluated based on the in vitro dissolution profilesin pH 7 phosphate buffer solution. Analysis of the effect of coatingthickness on LDEE release indicates that increasing the coating leveldecreases the in vitro release of active pharmaceutical ingredient (API)and although polymer has the sustained-release effect, its permeabilityis relatively large, thus a thick coating is required to prepareformulations with longer release duration (T90>5 hr).

In the final polymer coating formulation, talc was also added as alubricant to facilitate the fluid bed coating process at a ratio ofEudragit® E 100/talc at 10/1.

Development of Enteric Coating of Eudragit® E 100 Coated LDEE-S Beads

Initially, the enteric coating chosen at the development stage wasEudragit® S 100 and L 100 at a ratio of 2:1, and the ratio among polymerand other components was Eudragit® polymer: triethyl citrate (TEC):Talcratio of 7:2:1.

The in vitro dissolution profiles of prototype enteric coated beads(already coated with Eudragit® E 100 at a coating level of 65% w/w)coated with different levels of enteric film. The results showed that acoating level of 23% provides an adequate acid protection with less than5% LDEE released in acidic medium. Further, with less enteric coatinglevel (<10%), there is ˜20% LDEE released in acidic medium, and nosignificant difference in drug release profiles when coated at 5% or10%.

When the dissolution was done in pH 1 solution for 2 hr and then switchto pH 7 buffer, even with outer enteric coating layer, the permeabilityof inner Eudragit® E 100 layer may increase after 2 hr in pH 1.0 medium,since T90 was around 6.5 hr in pH 7 buffer for Eudragit® E coated beadsbut shortened to ˜4.5 hr in pH 7 buffer for enteric coated beads afterswitch over of dissolution medium.

For IPX203-B12-01, enteric polymer coatings that can dissolve at lowerpH were also developed, in which Eudragit® L 100-55 (dissolve above pH5.5) was used instead of Eudragit® S 100 and L 100. The ratio amongpolymer and other components in the coating formulation was Eudragit® L100-55:TEC:Talc of 6:1:3.

Dissolution Medium pH Effect on LDEE Release from Enteric Coated Beads

The effect of pH on the release of LDEE from the LDEE-S core beadscoated with Eudragit® E (65% w/w) and enteric coat (Eudragit® S 100/L100 at 2/1) was conducted at pH 1.0 solution for 2 hr and then switch topH 6.6, 6.8, 7.0 buffer solutions.

The results indicated that with less enteric layer coated (10%) orthinner enteric outer coating, drug release was earlier, and conversely,with a thicker enteric outer coating (23%), drug release was delayed atall pH's compared to the thinner enteric outer coated LDEE-S core beads.Further, with less or thinner enteric outer coating, there was no effecton drug release when pH changed from 6.6 to 6.8, and the drug releasewas slower when pH changed to 7.0. However, when thicker enteric coatlayer was applied (23%), there was no effect on drug release when pHchanged from 6.8 to 7.0, but the drug release was much slower when pHchanged to 6.6. Additionally, the pH value in dissolution medium canaffect drug release profiles through its effect on both enteric coatinglayer dissolution and Eudragit® E layer permeability. When enteric coatlayer is thin, its dissolution is fast and the pH effect on Eudragit® Eis more a rate-limiting factor. Since Eudragit® E permeability decreaseswith increasing pH, slower release was observed in pH 7.0 medium.However, with a thicker enteric coat, the dissolution of the entericlayer is much slower and become a rate-limiting step. With a combinationof Eudragit® S 100 and L 100 at a ratio of 2/1, its dissolution at lowerpH (pH 6.6) is much slower than at pH above 6.8. Thus the drug releaseis much slower in pH 6.6 medium with a thicker enteric coating.

Final Formulation of LDEE-S Beads for IPX203-B12-01

The test formulations for IPX203-B12-01 are summarized in Table 1. Thecomposition of the formulations of LDEE-S beads (IPX203-C0004,IPX203-C0005 and IPX203-C0006) is summarized in Table 2. FIG. 2 showsthe in vitro dissolution profiles of those formulations. IPX203-C0066was coated with 10% (w/w) enteric coat (Eudragit® S 100/L 100 at 2/1),which released ˜20% drug in the first 2 hr in pH 1.0 solution. Afterdissolution medium switch to pH 7 buffer, drug was controlled releasedover a period T90-3 hr. A better acidic protection for IPX203-0004 andIPX203-C0005 was observed due to their thicker enteric coat layer (25%w/w, Eudragit® L 100-55). Formulations IPX203-C0004 has a thinnerEudragit® E 100 layer of coating compared to IPX203-C0005, and has T90-3hr in pH 7 buffer. IPX203-C0005 provided longer release duration (T90˜5hr in pH 7 buffer).

TABLE 1 Test Formulations of IPX203 Prototype Capsule in Single DoseRelative Bioavailability (BA) Studies IPX203-B12-01*. Test LDEEFormulation Study (mg/2 capsules) IPX203-C0004 IPX203-B12-01 228IPX203-C0005 228 IPX203-C0006 228 *Carbidopa was dosed as commercialproduct Lodosyn ® 25 mg/tablet with the dosing regimen: 25 mg at T = 0and 6.25 mg (¼ tablet) at T = 4 hr.

TABLE 2 Composition of Final Formulation of LDEE-S Beads forIPX203-B12-01 Composition (w/w %) IPX203- IPX203- IPX203- IngredientC0004 C0005 C0006 Levodopa Ethyl Ester Succinate, 31.76 28.50 32.39Dihydrate Microcrystalline Cellulose, NF 14.66 13.15 14.95 Fumaric Acid,NF 2.44 2.19 2.49 Povidone, USP (Plasdone, K-29/32) 0.49 0.44 0.50 AminoMethacrylate Copolymer, NF 27.14 31.74 36.08 (Eudragit ® E 100)Methacrylic Acid Copolymer, Type 11.88 11.88 — C, NF (Eudragit ® L100-55) Methacrylic Acid Copolymer, Type — — 2.10 A, NF (Eudragit ® L100) Methacrylic Acid Copolymer, Type — — 4.20 B, NF (Eudragit ® S 100)Triethyl Citrate, NF 1.98 1.98 1.80 Talc, USP 9.64 10.12 5.50 Total100.0 100.0 100.0

II. In Vivo Results of IPX203-B12-01

The in vivo performance of the prepared formulations IPX203-C0004,IPX203-C0005 and IPX203-C0006 has been evaluated in healthy volunteersin a relative bioavailability analysis of IPX203-B12-01. The studydesign was a randomized, single-dose, crossover study in normal, healthyvolunteers under fasting condition.

FIG. 3 shows the plasma profile for the multi-particulate formulationsIPX203-C0004, IPX203-00005 and IPX203-C0006 in comparison with Sinemet®CR. All the IPX 203 multi-particulate formulations comprise Eudragit® Ecoating. The relative bioavailability parameters are provided in Table3. Comparison of the LD plasma concentration profile of the testedformulations to the reference product Sinemet® CR indicate that bothIPX203-C0005 and IPX203-C0006 showed sufficient AUC but more extendedeffect than Sinemet® CR, Further, the difference of Tmax betweenIPX203-C0004 and IPX203-C0005 corresponds well with their difference inin vitro dissolution profiles. Also, although the in vitro releaseprofiles for IPX203-C0004 and IPX203-00006 showed similar T90 (˜3 hr)after switch to pH 7 buffer, IPX203-C0006 showed more delayed effect invivo. Additionally, the results show that IPX203-C0006 has Cmax and AUCcomparable to those of Sinemet® CR.

TABLE 3 Relative LD Bioavailability Parameters of IPX203 Capsules Testedin Bioavailability Analysis of IPX203-B12-01 (n = 15). % Ratio DurationLD CD-LDEE of Test/ Concentra- Test (mg)^(a) Sinemet ® CR tion > 50%Formulation LDEE CD AUC₀-_(∞0)- Cmax Cmax (h)^(b) IPX203-C0004 228 31.2580 86 2.9 (3.3) IPX203-C0005 228 97  97′ 3.15 (3.25) IPX203-C0006 228 87104  3.25 (3.25) ^(a)LDEE 228 mg is equivalent to LD 200 mg.^(b)Sinemet ® CR tablet t_(max) = 2.5 hr

Example 2

I. Processing Procedures for Levodopa Ethyl Ester Succinate(LDEE-S)/Carbidopa (CD) Capsules for IPX203 B13-01

Preparation of Core Beads for IPX203-C0012, IPX203-C0013 andIPX203-C0016

Required amounts of LDEE-S-Dihydrate, Microcrystalline Cellulose,Fumaric acid, Povidone K29-32, ethanol and Purified Water weredispensed. The alcohol and the purified water were charged into acontainer and stirred using stir bar, Povidone was slowly added into theethanol/water mixed solvent. Mixing continued until the Povidone wascompletely dissolved, and the spray pump was calibrated to the targetgranulation spray rate.

LDEE-S-Dihydrate, Microcrystalline Cellulose, Fumaric acid, and Povidonewere charged into a high shear granulator and dry mixed for 1-5 minutesat impeller speed of 75 rpm and chopper speed of 1000 rpm. The Povidonesolution was sprayed into the granulation bowl and granulation continuedwith either ethanol or water as necessary. The granules were wet mixedfor 2 minutes, after the spraying was completed.

The wet granules were extruded using the extruder (MG 55 MultiGranulator) equipped with a 0.8 mm hole size screen at extruder speed of55 rpm. The extrudates were collected into double polyethylene-linedbags. The collected extrudate was weighed and adjusted in the quantitiesranging from 180-240 g per load.

One load of the weighed extrudate was charge into a spheronizer equippedwith a 3 mm cross hatch disc. The extrudate was spheronized at aspheronisation speed of 1400 rpm for 1-10 mins. The spheronized beadswere discharge into double PE bags. The remaining extrudate werespheronized until all the double polyethylene-lined bags are completed.

The wet beads were dried in a fluid bed drier (Glatt GPCP-1) at an Inlettemperature of 35±10° C. until Loss on Drying is not more than 5.0%. Thesteps above were repeated until additional sub loads have beenprocessed.

The dried beads were passed through a mechanical sieve (Vibroscreen)equipped with a 24-MG mesh screen at the bottom, 18-MG mesh screen inthe middle, and 16-MG mesh screen at the top. The beads that remained on18-US mesh and 24-MG mesh screens were collected into doublepolyethylene-lined bags.

Rate-Controlling Membrane Coating for IPX203-C0012 and IPX203-C0013

IPX203-C0012 Beads

Batch yield was determined. Based on the batch yield, the requiredamounts of Cellulose Acetate (CA) and Polyethylene Glycol 3350 (PEG3350)at weight ratio (CA/PEG) of 95/5 and Acetone/Purified Water (95/5 w/w)were calculated and dispensed. The Acetone was dispensed into astainless steel container and stirred using stir bar. While stirring,Cellulose Acetate (CA) was added slowly into the vortex of the solventand mixing was continued until the copolymer completely dissolved.

The Purified Water was dispensed into another stainless steel containerand was stirred using a stir bar. While stirring, Polyethylene Glycol3350 (PEG3350) was added slowly into the vortex of the purified watersolvent and mixing was continued until the copolymer completelydissolved. While stirring, PEG solution was added quickly into the CAsolution and mixing was continued until the solution was clear. Spraypump was calibrated to the target coating spray rate of the peristalticpump using the clear solution and the core beads were coated using GlattGPCG 1 equipped with a Wurster insert at Inlet air temperature of 33±10°C., Atomization air pressure of 1.0-2.0 bars and Wurster partitionheight of A0-40 mm. During coating, the inlet air temperature, exhaustflap, and spray rate were adjusted to maintain the exhaust airtemperature between 30±5° C.

After the target amount of coating solution was sprayed, the coatedbeads were dried at an inlet air temperature of 35±10° C. for 40-60minutes. The dried beads were passed through a mechanical sieve(Vibroscreen) equipped with a pan at the bottom and a 14-MG mesh screenat the top, and collected the beads that passed through the 14-MG meshscreen were collected in double polyethylene-lined bags. Oversized beadsthat remained on the 14-MG mesh screen were rejected.

IPX203-C0013 Beads

The procedure for preparing the coating solution and the coatingconditions are identical to those for IPX203-C0012 coating. However, therate-controlling polymer is Cellulose Acetate (CA), and the solvent isAcetone.

Muco-Adhesive Coating for IPX203-C0012, IPX203-C0013 and IPX203-C0016

The batch yield was determined. The required amounts of AminoMethacrylate Copolymer (Eudragit® E 100) and Talc were calculated anddispensed at weight ratio of 91/9. Purified Water, Acetone and IsopropylAlcohol were dispensed at weight ratio of 12/68/20 into a stainlesssteel container and stirred using stir bar. While stirring, AminoMethacrylate Copolymer (Eudragit® E 100) was slowly added into thevortex of the mixed solvent. Mixing continued until the copolymercompletely dissolved. While stirring, Talc was slowly dispersed into thevortex of the solution. Mixing continued until the material wascompletely dispersed. The suspension was continually stirred throughoutthe coating process.

The spray pump was calibrated to the target coating spray rate of theperistaltic pump using the suspension solution above. Therate-controlling membrane-coated beads for IPX203-C0012 andIPX203-C0013, or the core beads for IPX203-C0016 were coated with themuco-adhesive coating composition Glatt GPCG 1 equipped with a Wursterinsert at Inlet air temperature of 35±10° C., Atomization air pressureof 1.0-2.0 bars and Wurster partition height of 15-40 mm. Duringcoating, the inlet air temperature, exhaust flap, and spray rate wereadjusted to maintain the exhaust air temperature between 30±10° C.

After the target amount of coating solution was sprayed, the coatedbeads were dried at an inlet air temperature of 40±10° C. for 60-120minutes. The dried beads were passed through a mechanical sieve(Vibroscreen) equipped with a pan at the bottom and a 14-MG mesh screenat the top. The beads that passed through 14-MG mesh screen werecollected into double polyethylene-lined bags and the oversized beadsthat remained on the 14-MG mesh screen were rejected.

Enteric Coating for IPX203-C0012, IPX203-C0013 and IPX203-C0016

The batch yield was determined. Based on the batch yield, the requiredamounts of Triethyl Citrate, Talc and an enteric copolymer, eitherMethacrylic Acid Copolymer, Type A, (Eudragit L 100)/Methacrylic AcidCopolymer, Type B, (Eudragit® S) at ½ weight ratio for IPX203-C0012 andIPX203-C0016 or Eudragit® L 100 for IPX203-C00013 were calculated anddispensed. Acetone and Isopropyl Alcohol were dispensed at weight ratioof 40/60 into a stainless steel container and stirred using stir bar.While stirring, the enteric copolymer and Triethyl, Citrate (TEC) wereadded slowly into the vortex of the mixed solvent and mixing wascontinued until the enteric copolymer completely dissolved. Whilestirring, Talc was slowly dispensed into the vortex of the solution andmixing was continued until the material was completely dispersed. Thesuspension was continuously stirred throughout the coating process. Theweight ratio of enteric copolymer/TEC/Talc was 70/20/10.

The spray pump was calibrated to the target coating spray rate of theperistaltic pump using the solution, and the Eudragit® E-coated beadswere coated using Glatt GPCG 1 equipped with a Wurster insert at Inletair temperature of 35±10° C., Atomization air pressure of 1.0-2.0 barsand Wurster partition height of 15-30 mm. During coating, the inlet airtemperature, exhaust flap, and spray rate were adjusted to maintain theexhaust air temperature between 30±5° C. After the target amount ofcoating solution was sprayed, the coated beads were dried at an inletair temperature of 40±10° C. for 60-120 minutes and the dried beads werepassed through a mechanical sieve (Vibroscreen) equipped with a pan atthe bottom and a 14-MG mesh screen at the top. The beads that passedthrough 14-MG mesh screen were collected into double polyethylene-linedbags, and oversized beads that remained on the 14-MG mesh screen wererejected.

Immediate Release Granules (CD/LDEE-S)

The required amount of 27% Carbidopa USP, 49.9% Levodopa Ethyl EsterSuccinate-Dihydrate, 12.2% Dibasic Calcium Phosphate Anhydrous, 7.0%Hydroxypropyl Cellulose (Klucel-EXF), and 2.0% Croscarmellose Sodium,(Ac-Di-Sol) were dispensed and charged into the granulation bowl of ahigh shear granulator. The components were dry-mixed for 1-3 mins atimpeller speed between 150-250 rpm and Chopper Speed of 1000 rpm.Purified Water was sprayed at a desired flow rate into the granulationbowl until consistent wet mass was reached. The water/dry blend weightratio was between 0.20-0.40. The granules were wet-mixed for additional1-5 minutes, after the spraying was completed. The wet granules werecharged into the top spray product bowl of GPCG1 and dried using GPCG 1at inlet air temperature of 50° C. until the LOD is less than 6.0%.Inlet air flow was adjusted to maintain the fluidization of the wetgranules. The dried granules from the bowl were transferred into clean,double polyethylene-lined containers, and the granules were passedthrough the Fitzmill equipped with a stainless steel #24 mesh screen atKnife Mode and speed of 2000-3000 rpm. The required amount of Talc wascalculated based on the weight of the milled granules and 2% Talc of theimmediate release granules. The milled granules and Talc were chargedinto Pharmatech Miniblender and blended for 5 minutes. The blend wasdischarged into clean, double polyethylene-lined containers.

Encapsulation

The batch yield was determined. Based on the batch yield, the requiredamounts of the Enteric Coated Beads and Talc (at weight ratio of 99/1)were calculated and dispensed. The Enteric Coated Beads and Talc werecharged into an appropriated sized plastic bag and manually blended byshaking the plastic bag for 10 minutes. The blend and Immediate ReleaseGranules (CD/LDEE-S) were encapsulated with 00 size gelatin capsules,using MG Flexalab Encapsulator. For IPX203-C0016, the blend wasencapsulated but the Immediate Release Granules (CD/LDEE-S) were not.Table 4 shows the target fill weight for IPX203-C0012, IPX203-C0013 andIPX203-C0016 and Table 5 lists the composition of IPX203-C0012,IPX203-C0013 and IPX203-C0016.

TABLE 4 Target Fill Weight of IPX203-C0012, IPX203-C0013 andIPX203-C0016 Target Fill Weight (mg/Capsule) Enteric-coated BeadsImmediate release Granules IPX203-C0012 389.5 200.0 IPX203-C0013 412.0200.0 IPX203-C0016 252.6 N/A

TABLE 5 Formulation Composition of IPX203-C0012, IPX203-C0013 andIPX203-C0016 IPX203-C0012 1PX203-C0013 IPX203-C0016 Amount % Amount %Amount % Ingredient (mg/capsule) (w/w) (mg/capsule) (w/w) (mg/capsule)(w/w) Carbidopa, USP 54.0 9.2 54.0 8.8 Levodopa Ethyl Ester Succinate,306.4 52.0 306.4 50.1 81.8 32.4 Dihydrate Microcrystalline Cellulose, NF95.4 16.2 95.4 15.6 37.7 14.9 Amino Methacrylate Copolymer, 33.1 5.633.4 5.5 91.1 36.1 NF (Eudragit ® E 100) Fumaric Acid, NF (FineGranules) 15.9 2.7 15.9 2.6 6.3 2.5 Cellulose Acetate, NF (CA-398-10 9.11.5 12.9 2.1 0.0 NF) Talc, USP 13.1 2.2 15.2 2.5 13.9 5.5 MethacrylicAcid Copolymer, Type 8.5 1.4 10.6 4.2 B, NF (Eudragit ® S 100)Methacrylic Acid Copolymer, Type 4.3 0.7 25.9 4.2 5.3 2.1 A, NF(Eudragit ® L 100) Triethyl Citrate, NF 3.7 0.6 7.4 1.2 4.5 1.8Povidone, USP (Plasdone, K-29/32) 3.2 0.5 3.2 0.5 1.3 0.5 PolyethyleneGlycol, NF 0.5 0.1 Dibasic Calcium Phosphate, 24.3 4.1 24.3 4.0Anhydrous Hydroxypropyl Cellulose, NF 14.0 2.4 14.0 2.3 (Klucel-EXF)Croscarmellose Sodium, NF (Ac- 4.0 0.7 4.0 0.7 Di-Sol) Total 589.5 100.0612.0 100.0 252.6 100.0 *54 mg of Carbidopa, USP is equivalent to 50 mgof Carbidopa anhydrate. **306 mg of Levodopa Ethyl EsterSuccinate-Dihydrate is equivalent to 228 mg of Levodopa Ethyl Ester andto 200 mg Levodopa.II. Processing Procedures for Manufacturing Entacapone Capsules forIPX203 B13-01Preparation of Core Beads for IPX203-C0014 Capsule

The required amount of Entacapone, Microcrystalline Cellulose, PovidoneK29-32 and Purified Water were dispensed. The purified water was chargedinto a container and stirred using a stir bar, the Povidone (1.0% of thesolid blend) slowly added into the water at Povidone/Water weight ratioof 6/133.2 and mixing continued until the Povidone was completelydissolved. The spray pump was calibrated to the target granulation sprayrate (23 g/min), and 84.0% Entacapone and 15.0% MicrocrystallineCellulose were charged into a high shear granulator and were dry mixedfor 1-5 minutes at impeller speed of 200-300 rpm and chopper speed of1400-1600 rpm. The solution was sprayed into the granulation bowl untilall the solution was sprayed, and granulation was continued withPurified Water as necessary. The granules were wet-mixed for 2 minutes,after the spraying was completed. Then the wet granules were extrudedusing the extruder (MG 55 Multi Granulator) equipped with a 0.8 mm holesize screen at extruder speed of 50 rpm. The extrudates were collectedinto double polyethylene-lined bags. Further, the collected extrudatewere weighed and adjusted in the quantities ranging from 200-210 g perload.

One load of the weighed extrudate was charged into a spheronizerequipped with a 3 mm cross hatch disc and spheronized at spheronisationspeed of 1000 rpm for 1-2 mins. The spheronized beads were dischargedinto double PE bags. The wet beads were dried in a fluid bed drier(Glatt GPCP-1) at an Inlet temperature of 35±10° C. until Loss on Dryingwas not more than 5.0%. The dried beads were passed through a mechanicalsieve (Vibroscreen) equipped with a pan at the bottom, 24-MG mesh screenin the middle, and 16-MG mesh screen at the top. The beads that wereretained on 24-MG mesh were collected into double polyethylene-linedbags, and the beads on the pan and 16-MG mesh screen were rejected.

Enteric Coating for IPX203-C0014

The required amounts of Triethyl Citrate, Talc, Methacrylic AcidCopolymer Dispersion, NF (Eudragit® L 30D-55) and Water were calculatedand dispensed. The Purified Water was dispensed into a stainless steelcontainer and stirred using a stir bar. While stirring, Triethyl Citrate(TEC¹), Talc and the enteric copolymer dispersion were slowly added intothe vortex of Purified Water, and mixing was continued until thematerial was completely dispersed. The suspension was stirred throughoutthe coating process. The weight ratio of enteric copolymer/Talc/TEC was63.0/30.7/6.3.

The spray pump was calibrated to the target coating spray rate of theperistaltic pump using the solution, and the core beads were coatedusing Glatt GPCG 1 equipped with a Wurster insert at Inlet airtemperature of 35±10° C., Atomization air pressure of 1.0-2.0 bars andWurster partition height of 15-30 mm. During coating, the inlet airtemperature, exhaust flap, and spray rate were adjusted to maintain theexhaust air temperature between 30±5° C.

After the target amount of coating solution was sprayed, the coatedbeads were dried at an inlet air temperature of 30±10° C. until themoisture level was below 5%. The dried beads were passed through amechanical sieve (Vibroscreen) equipped with a pan at the bottom and a12-MG mesh screen at the top. The beads that passed through the 12-MGmesh screen were collected into double polyethylene-lined bags, and theoversized beads that remained on the 12-MG mesh screen were rejected.

Encapsulation for IPX203-C0014

The required amounts of the Enteric Coated Beads and Talc (at weightratio of 99/1) were calculated and dispensed, and the Enteric CoatedBeads and Talc were charged into an appropriate sized plastic bag. Thebeads and Talc were manually blended by shaking the plastic bag for atleast 5 minutes. The blend was encapsulated with 00 size gelatincapsules, using MG Flexalab Encapsulator. The target fill weight was 505mg. Table 6 lists the composition of IPX203-C0014.

TABLE 6 Formulation Composition of Entacapone Capsule (IPX203-C0014)Amount Ingredient % (w/w) (mg/capsule) Entacapone 79.2 400.0Microcrystalline Cellulose, NF 14.1 71.4 (Avicel PH-101) Povidone, USP(Plasdone, K-29/32) 1.0 4.8 Methacrylic Acid Copolymer Dispersion, 3.015.0 NF (Eudragit ® L 30D-55) Talc, USP 2.4 12.3 Triethyl Citrate, NF0.3 1.5 Total 100.0 505.0

III. In Vitro Release Profiles of Final LE(EE-S-Dihydrate Dosage Formsfor Pharmacokinetic (IPX203-B13-01)

Table 7 lists the test regimen for the 5-arm cross-over PK analysis(IPX203 B13-01).

TABLE 7 Dosing Regimen for IPX203 B13-01 CD/ LD/ Entacaponc/ RegimenDosage Form capsulc capsuIe capsule Regimen A IPX203-C0012 50 mg 200 mg*N/A Regimen B IPX203-C0012 + 50 mg 200 mg* 400 mg IPX203-C0014 Regimen CIPX203-C0013 + 50 mg 200 mg* 400 mg IPX203-C0014 Regimen DIPX203-C0013 + 50 mg 255.6 mg*  400 mg IPX203-C0016 + IPX203-C0014Regimen E Stalevo ® 37.5 mg  150 mg  200 mg 150 mg *LD equivalent dosebased on total amount of LDEE-S-Dihydrate in the formulation

The in vitro release profiles of the regimen A-D were measured using USPI dissolution method at agitation speed of 75 rpm in Simulated GastricFluid (pH 1.0) for first 2 hrs and followed by in Simulated IntestinalFluid (pH 7.0). FIG. 4 shows the release profiles of these testregimens.

The T90 (time duration for 90% of LDEE-S-Dihydrate released) isapproximately 3 hr, 4.5 h and 6 hrs for Regimen B, C and D,respectively. The LDEE-S-Dihydrate capsule (C0012) was used in bothRegimen A and Regimen B.

IV. In Vivo Evaluation (IPX203-B13-01)

The in vivo performance of the prepared dosage forms IPX203-C00012,IPX203-C00013 and IPX203-C00014 and IPX203-C0016 has been evaluated in12 healthy volunteers under fasted condition in a relativebioavailability analysis of IPX203-B13-01. The four test treatmentswere:

-   Regimen A: C0012-   Regimen B: C0012+C0014-   Regimen C: C0013+C0014

Regimen D: C0013+C0016+C0014

-   Regimen E: Stalevo 150 (Reference)    Where-   C0012 contained 228 mg LDEE ER beads with T90-3 hrs and 50 mg CD-   C0013 contained 228 mg LDEE ER beads with T90-5 hrs and 50 mg CD-   C0014 contained 400 mg enteric-coated entacapone-   C0016 contained 77 mg LDEE ER beads with T90-12 hrs

FIG. 5 shows the levodopa plasma profiles for all these regimens. Basedon the in vivo plasma profiles depicted in FIG. 5, the in vivo plasmaprofiles correlates well with the in vitro dissolution profiles depictedin FIG. 4. FIG. 5 demonstrates that Regimen D has the longesttherapeutic coverage and a constant plasma profile.

Example 3

Prepared Carbidopa Beads

The core beads of CD beads were formulated based on thegranulation-extrusion-spheronisation technology. 30 w/w % MCC was usedin the core seed formulation. No controlled release coating layer wasneeded. CD core beads was enteric-coated with the enteric coatingformulation comprising EUDRAGIT® S 100 and L 100 at a ratio of 2:1. Theenteric coating level was 5%.

Table 8 summarized the composition of final formulation of CD beads.

TABLE 8 Composition of Formulation of CD Beads Composition Ingredient(w/w %) Carbidopa 66.44 Microcrystalline Cellulose, NF 28.47 MethacrylicAcid Copolymer, Type A, NF 1.14 (Eudragit ® L 100) Methacrylic AcidCopolymer, Type B, NF 2.35 (Eudragit ® S 100)′ Triethyl Citrate, NF 1.00Talc, USP 0.60 Total 100.0

Example 4

The preparation procedure in Example 1 was repeated in this example,except the coating compositions. The core beads were coated first witheither cellulose acetate polymer or a combination of Hypromellose andethylcellulose. The coated beads were further coated with chitosan orpolycarbophil or Eudragit® E 100. After the second layer coating, thebeads were further coated with Eudragit® L 100-55. Table 9 shows thecomposition of four formulations IPX203-C0007, IPX203-C0008,IPX203-C0009 and IPX203-C0010.

TABLE 9 Composition of Formulations of LDEE-S Beads Using Chitosan orPolycarbophil as Muco-adhesive Polymers Composition (w/w %) IPX203-IPX203- IPX203- IPX203- Ingredient C0007 C0008 C0009 C0010 LevodopaEthyl Ester 39.45 45.18 31.14 45.30 Succinate, DihydrateMicrocrystalline 18.21 20.85 14.37 20.91 Cellulose, NF Fumaric Acid, NF3.03 3.48 2.40 3.48 Povidone, USP 0.61 0.70 0.48 0.70 (Plasdone,K-29/32) Hypromellose, Type 2.82 — 2.23 4.22 2910, USP (Pharmacoat 606,6 cps) Ethylcellulose, NF 11.28 — 8.90 16.89 (Ethocel, Standard-10 FPPremium) Polycarbophil, USP 3.77 — — 4.57 (Noveon ® AA-1) CelluloseAcetate, NF — 4.21 — — (CA-398-10 NF) Chitosan, NF — 3.74 — — (ChitoPharm ® S) (Material #50222178) Glacial Acetic Acid, USP — 1.01 — — AminoMethacrylate — — 17.86 — Copolymer, NF (Eudragit ® E 100) MethacrylicAcid 11.91 11.91 11.90 1.76 Copolymer, Type C, NF (Eudragit ® L 100-55)Triethyl Citrate, NF (PG) 1.99 1.99 1.98 0.29 Talc, USP 6.94 6.93 8.731.87 Total 100.0 100.0 100.0 100.0

Example 5

I. Formulations for IPX203-B14-01 Biostudy

Four test formulations were evaluated in biostudy IPX203-B14-01. ForIPX203-C0023, -C0024, and -C0025 formulations, there were two componentsin one capsule. For IPX203-C0026, there were three components in onecapsule. Table 10 below showed the formulation information for eachproduct, and Tables 11-13 showed formulation composition for eachcomponent.

TABLE 10 Test Formulations for Relative Bioavailability StudyIPX203-B14-01 Component Component Entacapone (ENT) I: IR II: LD ERComponent Test CD LD Prototype/ ENT Formulation (mg) (mg) LD (mg) (mg)IPX203-C0023 50 80 Prototype 0 1/280 1PX203-C0024 Prototype III/280IPX203-C0025 Prototype 11/280 IPX203-C0026 Prototype 200 11/280Stalevo ® 100 CD/LD/ENT (25/100/200 mg) (Reference)

TABLE 11 Composition of Prototype Formulations of IPX203 Component IIComposition (%) Ingredient Prototype I Prototype II Prototype III Corebead levodopa 65.26 62.15 61.03 Microcrystalline Cellulose 8.82 8.408.25 Mannitol, 8.82 8.40 8.25 Sodium Lauryl Sulfate 4.41 4.20 4.12Povidone 0.88 0.84 0.82 CA/Copovidone layer (1^(st) layer) CelluloseAcetate — 1.89 1.85 Copovidone — 2.31 2.27 Eudragit ® E 100 layer(2^(n<l) layer) Eudragit ® E 100 6.42 6.41 3.93 Talc 0.63 0.65 0.40Enteric layer (3^(rd) layer) Eudragit ® L 100 3.34 3.33 6.36 TriethylCitrate 0.96 0.95 1.81 Talc 0.47 0.48 0.91 Total 100.0 100.0 100.0

TABLE 12 Composition for Component I Formulation Ingredient Composition(w/w %) Carbidopa 35.86 Levodopa 53.14 Croscarmellose Sodium 7.00Povidone 3.00 Magnesium Stearate 1.00 Total 100.0

TABLE 13 Formulation Composition for Entacapone Component Ingredient %(w/w) Entacapone 73.15 Microcrystalline Cellulose, NF (Avicel 14.25PH-101) Povidone, USP (Plasdone, K-29/32) 1.90 Sodium Starch Glycolate3.80 Sodium Lauryl Sulfate, NF 1.90 Methacrylic Acid Copolymer, Type A,3.50 NF (Eudragit ® L 100) Talc, USP 0.50 Triethyl Citrate, NF 1.00Total 100.0II. Processing procedures for Manufacturing IPX203 Capsules for IPX203B14-01 BiostudyPreparation of Component I

Povidone was dissolved in the purified water completely, and then thespray pump with povidone solution was calibrated to the targetgranulation spray rate (40 mL/min). CD, LD, Croscarmellose Sodium werecharged into a high shear granulator and dry mixed for 1-5 minutes atimpeller speed of 150 rpm and chopper speed of 1800 rpm. While continuemixing, the solution from Step 1 was sprayed into the granulation bowluntil all the solution is sprayed, and granulation was continued withpurified water if necessary. The granules were collected, and the wetgranules were dried in a fluid bed drier (Glatt GPCP-1) at an Inlettemperature of 65° C. until Loss on Drying is not more than 2.5%. Thedried granules were passed through Fitzmill, and the material thatpasses through 30 mesh screen was collected. The collected material wasblended with magnesium stearate.

Alternative Preparation of Carbidopa-Containing Granules or Beads

In order to avoid potential carbidopa degradation during wet granulationprocess, a dry granulation process by roller compaction was developed.In this formulation, shown in Table 14, the procedures are described asbelow.

Appropriate amount of carbidopa, levodopa, microcrystalline cellulose,and croscarmellose sodium were charged into a suitable mixer. Thematerials were dry mixed for an appropriate time and then charged intoroller compactor at the controlled speed to start the roller compactionprocess. After roller compaction, the collected compacted sheets ofmaterials were blended with colloidal silicon dioxide for appropriatetime, and then milled into dried granules using a suitable mill. Finallythe milled granules were blended with magnesium stearate in the blender.

TABLE 14 Composition for Levodopa/Carbidopa IR Granules by DryGranulation Ingredient Composition (w/w %) Carbidopa 37.0 Levodopa 35.0Microcrystalline Cellulose 20.0 Croscarmellose Sodium 4.0 ColloidalSilicon Dioxide 3.0 Magnesium Stearate 1.0 Total 100.0

The amount and ratio of carbidopa and levodopa may be adjusted asdesired, so long as performance of the dried granules or beads are notcompromised.

Similarly, controlled release beads containing carbidopa may be preparedby a dry granulation method as provided through the incorporation ofrate-controlling excipient, muco-adhesive polymer, and/or enteric coat.Entacapone-containing beads or granules may also be prepared by a drygranulation method.

Preparation of Component II

Preparation of Core Beads for Component II

Povidone was dissolved in the purified water completely, and thencalibrate the spray pump with povidone solution to the targetgranulation spray rate (18 mL/min). LD, Microcrystalline Cellulose,Mannitol and Sodium Lauryl Sulfate were charged into a high sheargranulator and dry mixed for 1-5 minutes at impeller speed of 250 rpmand chopper speed of 1800 rpm. The solution from Step 1 was sprayed intothe granulation bowl until all the solution is sprayed, and granulationwith purified water was continued as necessary. The wet granules wereextruded using the extruder (MG 55 Multi Granulator) equipped with a 0.9mm hole size screen at extruder speed of 75 rpm. The extrudates werecollected, and the extrudates so collected were charged into aspheronizer equipped with a 3 mm cross hatch disc. The extrudates werespheronized at speed of 800 rpm for 1-2 mins. The wet beads were driedin a fluid bed drier (Glatt GPCP-1) at an Inlet temperature of 65±10° C.until Loss on Drying is not more than 2.5%. The dried beads were passedthrough a mechanical sieve (Vibroscreen) equipped with a 24-MG meshscreen at the bottom, and 16-MG mesh screen at the top. The beads thatremained on the 24-MG mesh screens were collected into doublepolyethylene-lined bags, and the oversized and undersized beads werediscarded.

CA/Copovidone Layer Coating for Component II

Cellulose acetate and copovidone (Kollidon VA64) were dissolved into themixture of acetone and isopropyl alcohol (IPA) solution (acetone/IPA atweight ratio of 4/1) completely. The pump was calibrated and set at thetarget spray rate of 15 g/min for the coating. The core beads from abovewere coated using Glatt GPCG 2 equipped with a Wurster insert at Inletair temperature of 35° C., Atomization air pressure of 2.0 bars andWurster partition height of 30 mm. During coating, the inlet airtemperature and spray rate were adjusted to maintain the exhaust airtemperature between 25±5° C. After the target amount of coating solutionwas sprayed, the coated beads were dried at an inlet air temperature of35° C. for 30 minutes. The dried beads were passed through a mechanicalsieve (Vibroscreen) equipped with a pan at the bottom and a 14-MG meshscreen at the top. The beads that passed through 14-MG mesh screen werecollected and the oversized beads were rejected.

Eudragit® E 100 Layer Coating

Acetone, IPA and purified water (at weight ratio acetone/IPA/water of68/20/12) were dispensed into a stainless steel container and beginstirring using stir bar. While stirring, Triethyl citrate, AminoMethacrylate Copolymer (Eudragit® E100) were slowly added into thevortex of the mixed solvent. The mixing was continued until thecopolymer completely dissolved. While stirring, Talc was slowlydispersed into the vortex of the solution. Mixing was continued untilthe material completely dispersed. The suspension was stirred throughoutthe coating process. The spray pump was calibrated to the target coatingspray rate (10 g/min) using the solution above. The beads (fromPrototype I, Prototype II and Prototype III) were coated using GlattGPCG 2 equipped with a Wurster insert at Inlet air temperature of 33°C., Atomization air pressure of 2.0 bars and Wurster partition height of30 mm. During coating, the inlet air temperature and spray rate wereadjusted to maintain the exhaust air temperature between 26±5° C. Afterthe target amount of coating solution was sprayed, the coated beads weredried at an inlet air temperature of 35° C. for 30 minutes. The driedbeads were passed through a mechanical sieve (Vibroscreen) equipped witha pan at the bottom and a 14-MG mesh screen at the top. The beads thatpassed through 14-MG mesh screen were collected, and the oversized beadswere rejected.

Enteric (Eudragit® L 100) Coating

Acetone and Isopropyl Alcohol were dispensed at weight ratio of 40/60into a stainless steel container and stirred using a stir bar. Whilestirring, the enteric copolymer Eudragit® L 100 and Triethyl Citrate(TEC) were slowly added into the vortex of the mixed solvent. Mixing wascontinued until the enteric copolymer completely dissolved. Whilestirring, Talc was slowly dispersed into the vortex of the solution.Mixing continued until the material was completely dispersed. Thesuspension was continually stirred throughout the coating process. Thespray pump was calibrated to the target at a coating spray rate (9g/min) using the solution above. The Eudragit® E-coated beads werecoated using Glatt GPCG 2 equipped with a Wurster insert at Inlet airtemperature of 35° C., Atomization air pressure of 2.0 bars and Wursterpartition height of 30 mm. During coating, the inlet air temperature,and spray rate was adjusted to maintain the exhaust air temperaturebetween 27±5° C. After the target amount of coating solution wassprayed, the coated beads were dried at an inlet air temperature of 38°C. for 30 minutes. The dried beads were passed through a mechanicalsieve (Vibroscreen) equipped with a pan at the bottom and a 14-MG meshscreen at the top. The beads that passed through 14-MG mesh screen werecollected, and the oversized beads were rejected.

Preparation of Entacapone Component (for IPX203-C0026)

Povidone was completely dissolved in the purified water. Entacapone,Sodium Starch Glycolate, Sodium Lauryl Sulfate and MicrocrystallineCellulose were charged into a high shear granulator and dry mix for 1-5minutes at impeller speed of 200-300 rpm and chopper speed of 1400-1600rpm. The solution was sprayed into the granulation bowl at the sprayrate of 19 ml/min until all the solution is used, and granulationcontinued with Purified Water as necessary. The wet granules wereextruded using the extruder (MG 55 Multi Granulator) equipped with a 0.9mm hole size screen at extruder speed of 55 rpm. The extrudates werecollected, and charged into a spheronizer equipped with a 3 mm crosshatch disc. The extrudate were spheronized at a spheronization speed of650 rpm for 2 mins. The wet beads were dried in a fluid bed drier (GlattGPCP-1) at an Inlet temperature of 40±5° C. until Loss on Drying is notmore than 5.0%. The dried beads were passed through a mechanical sieve(Vibroscreen) equipped with a pan at the bottom, 24-MG mesh screen inthe middle, and 16-MG mesh screen at the top. The beads that wereretained on 24-MG mesh were collected and the beads on the pan and 16-MGmesh screen were rejected. Acetone and Isopropyl Alcohol were dispensedat weight ratio of 40/60 into a stainless steel container and stirredusing a stir bar. While stirring, the enteric copolymer Eudragit® L 100and TEC were slowly added into the vortex of the mixed solvent. Mixingcontinued until the enteric copolymer completed dissolved. Whilestirring, Talc was slowly dispersed into the vortex of the solution.Mixing continued until the material was completely dispersed. Thesuspension was stirred throughout the coating process. The spray pumpwas calibrated to the target coating spray rate (8 g/min) using thesolution. The core beads were coated using Glatt GPCG 1 equipped with aWurster insert at Inlet air temperature of 35±10° C., Atomization airpressure of 1.5 bars and Wurster partition height of 15-30 mm. Duringcoating, the inlet air temperature, exhaust flap, and spray rate wereadjusted to maintain the exhaust air temperature between 27±5° C. Afterthe target amount of coating solution was sprayed, the coated beads weredried at an inlet air temperature of 40° C. for 20 minutes. The driedbeads were passed through a mechanical sieve (Vibroscreen) equipped witha pan at the bottom and a 14-MG mesh screen at the top. The beads thatpassed through the 14-MG mesh screen were collected and the oversizedbeads were rejected.

Encapsulation

The required amounts of the Component I and Component II Beads and Talcwere dispensed. For formulation IPX203-C0026, also Entacapone componentbeads were also dispensed. Talc was weighed at the weight ratio ofbeads/Talc at 99/1, and Component II beads and Talc were blendedthoroughly. For IPX203-C0026 product, Talc was also weighed at theweight ratio of ENT beads/Talc at 99/1, and Entacapone beads and Talcwere blended thoroughly. The Component I granules and Component II beads(from encapsulation section) were encapsulated into size 00 hard gelatincapsules, using MG Flexalab Encapsulator at the target fill weight forIPX203 products IPX203-C0023, -C0024, and -C0025. The Component Igranules, Component II beads (from encapsulation section), andentacapone beads (from encapsulation section) were encapsulated intosize 00 hard gelatin capsules, using MG Flexalab Encapsulator at thetarget fill weight for IPX203 products IPX203-C0026.

II. In Vitro LD Release Profiles of Four Formulations forPharmacokinetic Study (IPX203-B14-01)

The in vitro release profiles of IPX203-C0023, -C0024,-C0025 and -C0026were measured using USP 1 dissolution method at agitation speed of 75rpm in Simulated Gastric Fluid (pH 1.0, without enzyme) for first 2 hrsand followed by in Simulated Gastric Fluid (pH 7.0, without enzyme).FIG. 6 shows the release profiles of these four formulations.Formulation IPX203-C0025 and IPX203-C0026 contain the same Component IIbeads thus having same dissolution profiles. The T90 (time duration for90% of LD released) is approximately 4 hrs, 5 hrs and 7 hrs for -C0023,-C0025 and -C0025, and -C0026, respectively.

III. In Vivo Evaluation (Biostudy IPX203-B14-01)

The in vivo performance of the prepared products IPX203-C0023, -C0024,-C0025, and -C0026 has been evaluated in 19 healthy volunteers in arelative bioavailability study IPX203-B14-01. IPX203-B14-01 was asingle-center, open-label, randomized, single-dose, five-sequence,five-treatment crossover study. During each treatment period, subjectsreceived a single dose of the assigned study treatment. There was aminimum 5-day washout between treatments. Blood samples were obtainedpredose and following dosing for approximately 12 hours for measurementof plasma concentrations. Thirty healthy male and female subjects, 18 to45 years of age at the time of dosing with a body mass index of 18.0 to30.0 kg/m², inclusive, were enrolled. All treatments were administeredwith 240 mL of room-temperature water to subjects in a fasted state.Subjects were instructed to swallow the study drugs intact withoutcrushing or chewing. FIG. 7 shows the levodopa plasma profiles for allthese regimens, and Table 15 shows the PK parameters relative toStalevo®.

TABLE 15 PK Parameters for All the Regimens Tested in IPX203 B13-01Study (n = 19) % of IPX203 Test % of IPX203 Test Formulation/Stalevo ®Formulation/Stalevo ® (normalized by LD dose) Formulation AUC₀-_(∞) CmaxAUC₀-_(∞) Cmax IPX203-C0023 277.3 179.4 77.0 49.8 IPX203-C0024 199.1121.4 55.3 33.7 IPX203-C0025 226.9 134.0 63.0 37.2 IPX203-C0026 265.9141.6 73.9 39.3

Table 16 shows the duration of time above 50% Cmax for IPX203-C0023,-C0024, -C0025, and -C0026 and conventional formulations.

TABLE 16 Duration of Time Above 50% Cmax IPX203-C0023, -C0024, -C0025and -C0026 and Conventional Formulations % Coefficient of Formulation NMedian Mean Variation (SD/Mean) IPX203-C0023 19 4.00 4.14 29.88IPX203-C0024 19 5.38 4.84 35.94 IPX203-C0025 19 5.38 5.20 29.30IPX203-C0026 19 4.88 5.23 36.32 C_(max) Values normalized to allowcomparison

Comparison of the LD plasma concentration profile of the testedformulations to the reference product Stalevo® indicates that: (1) theIPX203 regimens, based on IPX203-C0023, -C0024, -C0025, and -C0026formulations, showed more extended effect than Stalevo® (Table 16 andFIG. 7); in addition, the IPX203 formulations showed more extendedeffect than Sinemet® or Sinemet® CR (Table 16 and FIG. 3; for Sinemet®CR (N=11), T>50% Cmax is ˜3.41 hrs); (2) the IPX203 formulations; namelyIPX203-C0023, -C0024, -C0025, and -C0026 formulations, showed relativelyflat plasma profiles for LD compared to Stalevo® (FIG. 7); (3) the timeduration between 50% of Cmax to Cmax for IPX203-C0023, -C0024, -C0025,and -C0026 formulations are much longer than Stalevo®, (approximately4.1-5.2 hrs for test formulations, compared to 2.3 hrs for Stalevo®);and (4) the variation of the time duration between 50% Cmax to Cmax forIPX203-C0023, -C0024, -C0025, and -C0026 formulations is less thanStalevo®.

The invention illustratively described herein suitably may be practicedin the absence of any element or elements, limitation or limitationswhich is not specifically disclosed herein. Thus, for example, in eachinstance herein, any of the terms “comprising,” “consisting essentiallyof” and “consisting of” may be replaced with either of the other twoterms. The terms and expressions which have been employed are used asterms of description and not of limitation, and there is no intention inthe use of such terms and expressions of excluding any equivalents ofthe features shown and described or portions thereof, but it isrecognized that various modifications are possible within the scope ofthe invention claimed. Thus, it should be understood that although thepresent invention has been specifically disclosed by preferredembodiments and optional features, modification and variation of theconcepts herein disclosed may be resorted to by those skilled in theart, and that such modifications and variations are considered to bewithin the scope of this invention as defined by the appended claims.

What is claimed is:
 1. A method for treating Parkinson's disease orprimary parkinsonism comprising orally administering a multiparticulateformulation to a human patient three or four times a day wherein themultiparticulate formulation comprises: (a) a plurality of controlledrelease particles wherein the controlled release particles aremini-tablets, beads or granules and comprise: i) a core comprisinglevodopa, optionally carbidopa, and at least one pharmaceuticallyacceptable excipient; ii) a muco-adhesive layer or coating surroundingthe core wherein the muco-adhesive layer comprises an amino methacrylatecopolymer; and iii) an enteric layer or coating surrounding themuco-adhesive layer or coating wherein the enteric layer or coatingcomprises at least one enteric polymer; (b) an immediate releasecomponent comprising levodopa and optionally carbidopa and wherein theimmediate release component is a powder, mini-tablet, bead, pellet,granule, a coating applied to the controlled release particles, or acombination thereof.
 2. The method of claim 1 wherein the controlledrelease particles release: 0-60% of the levodopa after 2 hours; 25-90%of the levodopa after 4 hours; and 35-100% of the levodopa after 6 hourswhen the controlled release particles are tested using a United StatesPharmacopeia (USP) Type I or USP Type II dissolution apparatus at 37°C., with a rotational speed of 75 rpms and 900 mL of an aqueous mediawith a pH of
 7. 3. The method of claim 1 wherein the multiparticulateformulation releases at least 90% of the levodopa in approximately 5 to7 hours when tested in a United States Pharmacopeia (USP) Type Idissolution apparatus at 37° C. with a rotational speed of 75 rpms and900 mL of simulated gastric fluid with a pH of 1.0 for 2 hours followedby simulated intestinal fluid with a pH of 7.0.
 4. The method of claim 1wherein the muco-adhesive layer further comprise a polymer selected fromthe group consisting of polycarbophil, carbomer, cellulosics, chitosan,diethylaminodextran, diethylaminoethyldextran, polygalactosamine,polylysine, polyomithine, prolamine, polyimine, hyaluronic acid, sodiumalginate, sodium carboxymethylcellulose and alginate or a combinationthereof.
 5. The method of claim 1, wherein the amino methacrylatecopolymer is a dimethylaminoethyl methacrylate copolymer.
 6. The methodof claim 1, wherein the enteric polymer is selected from the groupconsisting of shellac, cellulose acetate phthalate, methacrylic acidcopolymers, cellulose acetate trimellitate, poly(vinyl acetatephthalate), hydroxypropyl methylcellulose phthalate and hydroxypropylmethylcellulose acetate succinate.
 7. The method of claim 6 wherein theenteric polymer is a methacrylic acid copolymer.
 8. The method of claim1 wherein the controlled release particles have a size that passesthrough a 12 mesh screen but are retained on a 25 mesh screen.
 9. Themethod of claim 1 wherein the controlled release particles comprises:(i) the core comprising levodopa and at least one pharmaceuticallyacceptable excipient; (ii) a first layer surrounding the core comprisingat least one rate-controlling polymer; (iii) the muco-adhesive layersurrounding the first layer; and (iv) the enteric layer surrounding themuco-adhesive layer comprising at least one enteric polymer.
 10. Amethod for treating Parkinson's disease or primary parkinsonismcomprising orally administering a multiparticulate formulation to ahuman patient three or four times a day wherein the multiparticulateformulation comprises: (a) a plurality of controlled release particleswherein the controlled release particles are mini-tablets, beads orgranules and comprise: (i) a core comprising levodopa, optionallycarbidopa, and at least one pharmaceutically acceptable excipient; (ii)a first layer surrounding the core comprising at least onerate-controlling polymer; (iii) a second layer surrounding the firstlayer comprising an amino methacrylate copolymer and optionally one ormore additional muco-adhesive polymers selected from the groupconsisting of polycarbophil, carbomer, cellulosic, chitosan,diethylaminodextran, diethylaminoethyldextran, polygalactosamine,polylysine, polyomithine, prolamine, polyimine, hyaluronic acid, sodiumalginate, sodium carboxymethylcellulose and alginate or a combinationthereof; and (iv) a third layer surrounding the second layer comprisingat least one enteric polymer selected from the group consisting ofshellac, cellulose acetate phthalate, methacrylic acid copolymers,cellulose acetate trimellitate, poly(vinyl acetate phthalate),hydroxypropyl methylcellulose phthalate and hydroxypropylmethylcellulose acetate succinate; and (b) an immediate releasecomponent comprising levodopa and optionally carbidopa wherein theimmediate release component is a powder, mini-tablet, bead, pellet,granule, a coating applied to the controlled release particles, or acombination thereof.
 11. The method of claim 10 wherein the controlledrelease particles pass through a 14 mesh screen but are retained on a 24mesh screen.
 12. The method of claim 10 wherein the controlled releaseparticles release: 10-55% of the levodopa after 2 hours; 30-85% of thelevodopa after 4 hours; and 40-100% of the levodopa after 6 hours whenthe controlled release particles are tested using a United StatesPharmacopeia (USP) Type I or USP Type II dissolution apparatus at 37°C., with a rotational speed of 75 rpms and 900 mL of an aqueous mediawith a pH of
 7. 13. The method of claim 10 wherein the multiparticulateformulation releases at least 90% of the levodopa in approximately 5 to7 hours when tested in a USP Type I dissolution apparatus at 37° C. witha rotational speed of 75 rpms and 900 mL of simulated gastric fluid witha pH of 1.0 for 2 hours followed by simulated intestinal fluid with a pHof 7.0.
 14. A multiparticulate formulation comprising: (a) a pluralityof controlled release particles wherein the controlled release particlesare mini-tablets, beads, or granules comprising: i) a core comprisinglevodopa, optionally carbidopa, and at least one pharmaceuticallyacceptable excipient; ii) a muco-adhesive layer or coating surroundingthe core wherein the muco-adhesive layer comprises an amino methacrylatecopolymer; and iii) an enteric layer or coating surrounding themuco-adhesive layer or coating wherein the enteric layer or coatingcomprises at least one enteric polymer; (b) an immediate releasecomponent comprising levodopa and optionally carbidopa wherein theimmediate release component is a powder, mini-tablet, bead, pellet,granule, a coating applied to the controlled release particles, or acombination thereof.
 15. The formulation of claim 14 wherein thecontrolled release particles release: 0-60% of the levodopa after 2hours; 25-90% of the levodopa after 4 hours; and 35-100% of the levodopaafter 6 hours when the controlled release particles are tested using aUnited States Pharmacopeia (USP) Type I or USP Type II dissolutionapparatus at 37° C., with a rotational speed of 75 rpms and 900 mL of anaqueous media with a pH of
 7. 16. The formulation of claim 14 whereinthe formulation releases at least 90% of the levodopa in approximately 5to 7 hours when tested in a United States Pharmacopeia (USP) Type Idissolution apparatus at 37° C. with a rotational speed of 75 rpms and900 mL of simulated gastric fluid with a pH of 1.0 for 2 hours followedby simulated intestinal fluid with a pH of 7.0.
 17. The formulation ofclaim 14 wherein the muco-adhesive layer further comprise a polymerselected from the group consisting of polycarbophil, carbomer,cellulosics, chitosan, diethylaminodextran, diethylaminoethyldextran,polygalactosamine, polylysine, polyomithine, prolamine, polyimine,hyaluronic acid, sodium alginate, sodium carboxymethylcellulose andalginate or a combination thereof.
 18. The formulation of claim 14,wherein the amino methacrylate copolymer is a dimethylaminoethylmethacrylate copolymer.
 19. The formulation of claim 14, wherein theenteric polymer is selected from the group consisting of shellac,cellulose acetate phthalate, methacrylic acid copolymers, celluloseacetate trimellitate, poly(vinyl acetate phthalate), hydroxypropylmethylcellulose phthalate and hydroxypropyl methylcellulose acetatesuccinate.
 20. The formulation of claim 19 wherein the enteric polymeris a methacrylic acid copolymer.
 21. The formulation of claim 14 whereinthe controlled release particles have a size that passes through a 12mesh screen but are retained on a 25 mesh screen.
 22. The formulation ofclaim 14 wherein the controlled release particles comprises: (i) thecore comprising levodopa and at least one pharmaceutically acceptableexcipient; (ii) a first layer surrounding the core comprising at leastone rate-controlling polymer; (iii) the muco-adhesive layer surroundingthe first layer; and (iv) the enteric layer surrounding themuco-adhesive layer comprising at least one enteric polymer.
 23. Theformulation of claim 14 wherein the formulation produces an in vivolevodopa plasma profile following oral administration of the formulationto a human subject under fasting conditions comprising: (a) a levodopaplasma concentration corresponding to maximum levodopa plasmaconcentration (C_(max)) occurring within 6 hours after administration ofthe formulation; (b) a time to reach 50% C_(max) of less than one hourafter administration of the formulation; and (c) wherein the in vivoplasma level of levodopa is maintained at 50% C_(max) or above for atleast 5.0 hours after administration of the formulation.
 24. Theformulation of claim 14 wherein the controlled release particlescomprise: (i) a core comprising levodopa, optionally carbidopa, and atleast one pharmaceutically acceptable excipient; (ii) a first layersurrounding the core comprising at least one rate-controlling polymer;(iii) a second layer surrounding the first layer comprising an aminomethacrylate copolymer and optionally one or more additionalmuco-adhesive polymers selected from the group consisting ofpolycarbophil, carbomer, cellulosic, chitosan, diethylaminodextran,diethylaminoethyldextran, polygalactosamine, polylysine, polyomithine,prolamine, polyimine, hyaluronic acid, sodium alginate, sodiumcarboxymethylcellulose and alginate or a combination thereof; and (iv) athird layer surrounding the second layer comprising at least one entericpolymer selected from the group consisting of shellac, cellulose acetatephthalate, methacrylic acid copolymers, cellulose acetate trimellitate,poly(vinyl acetate phthalate), hydroxypropyl methylcellulose phthalateand hydroxypropyl methylcellulose acetate succinate.
 25. The formulationof claim 24 wherein the controlled release particles pass through a 14mesh screen but are retained on a 24 mesh screen.
 26. The formulation ofclaim 24 wherein the controlled release particles release: 10-55% of thelevodopa after 2 hours; 30-85% of the levodopa after 4 hours; and40-100% of the levodopa after 6 hours when the controlled releaseparticles are tested using a United States Pharmacopeia (USP) Type I orUSP Type II dissolution apparatus at 37° C., with a rotational speed of75 rpms and 900 mL of an aqueous media with a pH of
 7. 27. Theformulation of claim 24 wherein the formulation releases at least 90% ofthe levodopa in approximately 5 to 7 hours when tested in a USP Type Idissolution apparatus at 37° C. with a rotational speed of 75 rpms and900 mL of simulated gastric fluid with a pH of 1.0 for 2 hours followedby simulated intestinal fluid with a pH of 7.0.